Nitrogen-containing benzoheterocycle compound comprising carboxylic acid group, preparation method and use thereof

ABSTRACT

Related is a nitrogen-containing benzoheterocycle compound containing a carboxylic acid group as shown in general formula (I), or a stereoisomer or pharmaceutically acceptable salt thereof. The compound has excellent pharmacokinetic properties and extremely remarkable liver targeting properties, and a pharmaceutical composition comprising the same may be used as a CCR antagonist, in particular a CCR2 and/or CCR5 antagonist and can be used in mediated disease, including, but not limited to, nonalcoholic fatty liver disease (NAFLD) or the like.

TECHNICAL FIELD

The present application relates to a nitrogen-containingbenzoheterocycle compound containing a carboxylic acid group as shown ingeneral formula (I), or a stereoisomer or pharmaceutically acceptablesalt thereof, and a pharmaceutical composition comprising the same. Thecompound has excellent pharmacokinetic properties and extremelyremarkable liver targeting properties, and can be used as a CCRantagonist, in particular a CCR2 and/or CCR5 antagonist. Therefore, thepresent application further relates to use of the compound in preventionand/or treatment of a disease mediated by CCR (in particular CCR2 and/orCCR5), including, but not limited to, nonalcoholic fatty liver disease(NAFLD).

BACKGROUND ART

The nonalcoholic fatty liver disease (NAFLD) refers to accumulation ofexcess fat in the form of triglycerides (steatosis) in the liver. It isdefined to include, first of all, the presence of hepatic steatosisverified by liver imaging or histology, and secondly the absence ofother factors that lead to secondary hepatic steatosis, such as alcoholconsumption, drugs, genetic abnormalities and the like. The diseasespectrum includes nonalcoholic simple fatty liver (NAFL), nonalcoholicsteatohepatitis (NASH) and related cirrhosis and liver cancer.

The exact pathogenesis of NASH has not been elucidated and it is almostdifferent in each patient. Although the disease is closely interrelatedwith insulin resistance, obesity, and metabolic syndrome, not allpatients in these states will suffer from NAFLD and(or) NASH, and notall patients suffering from NAFLD and(or) NASH will experience thesestates. If NAFLD develops into NASH, risks of occurrences of cirrhosis,liver failure and liver cancer will increase greatly.

As researches on the pathogenesis of the nonalcoholic steatohepatitis(NASH) continue, looking for some new therapeutic targets or drugsbecomes hotspots at now. The mechanisms of new drug against NASH aremainly classified into four categories: diabetes/lipid, bile acid/bileacid receptor pathway, apoptosis, and anti-inflammation/fibrosis. Themain targets for NASH include peroxisome proliferator-activatedreceptor-α (PPARA) and β (PPARD) agonists, farnesyl ester-X receptor(FXR) agonists, monoclonal antibodies of inhibiting lysyl oxidase-like 2(LOXL2) proteins, apoptosis signal-regulating protein kinase (ASK1)inhibitors, acetyl-CoA carboxylase (ACC) protein allosteric inhibitors,and chemokine receptor (CCR2/CCR5) antagonists.

Currently, drugs for NASH are not yet commercially available, and allcompounds are still in clinical research stage. Obeticholic acid fromIntercept Co. is the fastest-moving drug, and which is in phase IIIclinical studies. For Ceniciviroc from Takeda Pharmaceutical Co. ofJapan phase-II clinical studies have been completed. Internationalpharmaceutical companies such as Gilead, Bristol-Myers Squibb, and AstraZeneca all have compounds in phase-II clinical studies. Medical therapyfor NASH has become a research hotspot in the global medical field.WO03014105 describes a bicyclic compound and its use in anti-HIV,wherein R¹ is C₁₋₆ alkoxy-C₁₋₆ alkoxy; R² is tetrahydropyran-amino,alkyl-substituted imidazolyl, and alkyl-substituted triazolyl; R³ ishalogen, substituted alkyl, substituted alkoxy; Y^(a) is oxygen,sulfinyl, sulfuryl, alkyl-substituted amino, and Z^(2a) is sulfur,sulfinyl, sulfuryl. The specific description in this patent are notconsidered as part of the invention, and the compound therein has thefollowing formula:

EP1593681 describes a tricyclic compound and use thereof in anti-HIV,wherein R^(1a) is C₁₋₆ alkoxy-C₁₋₆ alkoxy; R^(2a) istetrahydropyran-amino, alkyl-substituted imidazolyl, andalkyl-substituted triazolyl; and Z^(2a) may be sulfur, sulfinyl,sulfuryl; W is a group of formula (a) or (b); R³ is halogen, substitutedalkyl, substituted alkoxy. The specific description in this patent arenot considered as part of the invention, and the compound therein hasthe following formula:

WO03076411 describes methods for synthesis of optical isomers havingformula (c) and medical uses thereof for the treatment of HIV. Thespecific description in this patent is not considered as part of theinvention, and the compound therein has the following formula:

WO2015143367 describes uses of Cenicriviroc for the treatment of HIV andfibrosis, and WO2016105527 describes methods for the synthesis ofCenicriviroc. The specific description in these patents are notconsidered as part of the invention.

SUMMARY OF THE INVENTION

Through deep researches, the inventor surprisingly found that thecompound obtained by introducing a carboxylic acid group into anitrogen-containing benzoheterocyclic compound has very potentinhibitory effect against CCR2/CCR5, but has no obvious inhibitoryeffect against CYP and hERG, and has good safety, as well as extremelyremarkable liver targeting properties and excellent pharmacokineticproperties. The compound of the present application may be used for thetreatment of a CCR2- and/or CCR5-mediated disease, including, but notlimited to, nonalcoholic fatty liver disease (NAFLD) or the like. Thepresent application is accomplished based on the above.

Particularly, the present application relates to the following:

In a first aspect, the present application provides a compound ofgeneral formula I or a pharmaceutically acceptable salt, ester, solvate(e.g., hydrate), stereoisomer, tautomer, prodrug thereof, or crystallineform, metabolite thereof, or a mixture of the aforementioned:

wherein R¹ is selected from hydrogen, deuterium, and C₁₋₆ alkyl, saidC₁₋₆ alkyl is optionally substituted with R²;

L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene,C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene;

L₂ is absent or selected from —NH—, C₂₋₆ alkylene, C₂₋₆ alkenylene,—(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene,C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene and 3- to 10-memberedheterocyclylene; said —NH—, C₂₋₆ alkylene, C₂₋₆ alkenylene, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and 3- to 10-memberedheterocyclylene are optionally substituted with one or more substituentsindependently selected from hydrogen, deuterium, halogen, hydroxyl,cyano, C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy,—C(O)C₁₋₄ alkyl, —COOR³, —N(R³)₂, C₃₋₁₀ cycloalkyl, 3- to 10-memberedheterocyclic group, C₆₋₁₀ aryl, —CON(R³)₂ and —NR³CO₂R³; said 3- to10-membered heterocyclylene contains one or more ring members selectedfrom N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2;

L³ is absent or selected from C₁₋₆ alkylene, C₂₋₆ alkenylene, C₂₋₆alkynylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and3- to 10-membered heterocyclylene;

R² is selected from hydrogen, deuterium, hydroxyl, amino, carboxyl, C₁₋₆alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆ alkyleneoxy)_(m2)-C₁₋₆ alkyl,(C₁₋₆ alkyl)₂ amino, and 5- to 6-membered heterocyclyl containing one ormore N, O or S; wherein m₁, m₂ are each independently selected from 0,1, 2, 3 or 4;

R³ is selected from hydrogen, deuterium, C₁₋₄ alkyl and C₃₋₆ cycloalkyl;

A is selected from 4- to 7-membered heterocyclyl or 5- to 10-memberedheteroaryl, both of which are optionally substituted with a substituentselected from deuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl that areoptionally substituted with R⁶, halogen, —CN, halogenated C₁₋₆ alkyl,C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂ heteroaryl; R⁶ is selectedfrom deuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆ cycloalkoxy;

n is selected from 0, 1 or 2.

In some preferred embodiments, R¹ is selected from hydrogen and C₁₋₄alkyl, said C₁₋₄ alkyl is optionally substituted with R²; R² is selectedfrom hydroxyl, carboxyl, C₁₋₆ alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆alkyleneoxy)_(m2)-C₁₋₆ alkyl and 5- to 6-membered heterocyclylcontaining one or more N, O or S; wherein m₁ and m₂ are independentlyselected from 0, 1, 2, 3 or 4.

In some preferred embodiments, R¹ is selected from C₁₋₄ alkyl, said C₁₋₄alkyl is optionally substituted with R²; R² is selected from hydroxyl,C₁₋₆ alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆ alkyleneoxy)_(m2)-C₁₋₆ alkyland 5- to 6-membered heterocyclyl containing one or more N, O or S;wherein m₁ and m₂ are each independently selected from 0, 1 or 2.

In some preferred embodiments, R¹ is selected from C₁₋₄ alkyl, said C₁₋₄alkyl is optionally substituted with R²; R² is selected from C₁₋₄alkoxy, —(C₁₋₄ alkyleneoxy)_(m1)-(C₁₋₄ alkyleneoxy)_(m2)-C₁₋₄ alkyl and5- to 6-membered heterocyclyl containing one or more N, O or S; whereinm₁ and m₂ are each independently selected from 0, 1 or 2.

In some preferred embodiments, R¹ is selected from C₁₋₄ alkyl, said C₁₋₄alkyl is optionally substituted with R²; R² is selected from C₁₋₄alkoxy.

In some preferred embodiments, R¹ is

In some preferred embodiments, L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, and C₅₋₁₂ heteroarylene.

In some preferred embodiments, L₁ is selected from C₁₋₆ alkylene, C₃₋₇cycloalkylene, C₆₋₈ arylene, and C₅₋₈ heteroarylene.

In some preferred embodiments, L₁ is selected from C₁₋₃ alkylene andC₃₋₅ cycloalkylene.

In some preferred embodiments, L₁ is selected from C₁₋₃ alkylene.

In some preferred embodiments, L₁ is selected from methylene andethylene.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₆alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and 3- to 10-memberedheterocyclylene; said —NH—, C₂₋₆ alkenylene, —(C₁₋₆alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆-10arylene, C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene areoptionally substituted with one or more substituents independentlyselected from hydrogen, deuterium, halogen, hydroxyl, cyano, C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, —C(O)C₁₋₄ alkyl, —COORS,—N(R³)₂, C₃₋₁₀ cycloalkyl, 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl,—CON(R³)₂ and —NR³CO₂R³; said 3- to 10-membered heterocyclylene containsone or more ring members selected from N, NR³, O, P, and S(O)_(z);wherein y₁ and y₂ are each independently selected from 0, 1, 2, 3 or 4;z is selected from 0, 1 or 2.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, C₁₋₃ alkyl, C₁₋₃haloalkyl, C₁₋₃ haloalkoxy, —C(O)C₁₋₄ alkyl, —COORS and —NR³CO₂R³; said3- to 10-membered heterocyclylene contains one or more ring membersselected from N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2; R³ is selected from hydrogen, deuterium, C₁₋₄ alkyl and C₃₋₆cycloalkyl.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, methyl, halomethyl,halomethoxy, —C(O)CH₃, —COOR³ and —NR³CO₂R³; said 3- to 10-memberedheterocyclylene contains one or more ring members selected from N, NR³,O, P, and S(O)_(z); wherein y₁ and y₂ are each independently selectedfrom 0, 1, 2, 3 or 4; z is selected from 0, 1 or 2; R³ is selected fromhydrogen, deuterium, methyl and cyclopropyl.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and 3- to 10-memberedheterocyclylene are optionally substituted with one or more substituentsindependently selected from hydrogen, deuterium, halogen, hydroxyl,cyano, methyl, halomethyl, halomethoxy, —C(O)CH₃, —COOR³ and —NR³CO₂R³;said 3- to 10-membered heterocyclylene contains one or more ring membersselected from N, NR³, O, P, and S(O)_(z); wherein y₁, y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2; R³ is selected from hydrogen, deuterium, methyl and cyclopropyl.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₄alkylene, C₂₋₃ alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 6-membered heterocyclylene; said —NH—, C₂₋₄alkylene, C₂₋₃ alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 6-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, methyl, halomethyl,halomethoxy, —C(O)CH₃, —COOR³ and —NR³CO₂R³; said 3- to 6-memberedheterocyclylene contains one or more ring members selected from N, NR³,O, P, and S(O)_(z); wherein y₁, y₂ are each independently selected from0, 1, 2, 3 or 4; z is selected from 0, 1 or 2; R³ is selected fromhydrogen, deuterium, methyl and cyclopropyl.

In some preferred embodiments, L₂ is absent or selected from —NH—, C₂₋₄alkylene, C₂₋₃ alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 6-membered heterocyclylene; said —NH—, C₂₋₃alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene, and 3- to 6-memberedheterocyclylene are optionally substituted with one or more substituentsindependently selected from hydrogen, deuterium, halogen, hydroxyl,cyano, methyl, halomethyl, halomethoxy, —C(O)CH₃, —COORS and —NR³CO₂R³;said 3- to 6-membered heterocyclylene contains one or more ring membersselected from N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2; R³ is selected from hydrogen, deuterium, methyl and cyclopropyl.

In some preferred embodiments, L₂ is absent or selected from —NH—,—N(CH₃)—, —C(CH₃)₂—, —CH═CH—, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 6-membered heterocyclylene, wherein y₁ and y₂are each independently selected from 0, 1, or 2.

In some preferred embodiments, L₂ is absent.

In some preferred embodiments, L₂ is selected from C₂₋₄ alkylene.

In some preferred embodiments, L₂ is selected from —NH—, —N(CH₃)— and—C(CH₃)₂—.

In some preferred embodiments, L₂ is —C(CH₃)₂—.

In some preferred embodiments, L₃ is absent or selected from C₁₋₃alkylene, C₂₋₃ alkenylene, C₂₋₃ alkynylene, C₃₋₆ cycloalkylene, C₆₋₈arylene, C₅₋₈ heteroarylene, and 3- to 7-membered heterocyclylene.

In some preferred embodiments, L₃ is absent or selected from C₁₋₆alkylene and C₂₋₆ alkenylene.

In some preferred embodiments, L₃ is absent or selected from —C(CH₃)₂—,—CH₂—, and —CH═CH—.

In some preferred embodiments, L₃ is absent.

In some preferred embodiments, L₃ is —CH₂—.

In some preferred embodiments, A is selected from 4- to 7-memberedheterocyclyl or 5- to 10-membered heteroaryl.

In some preferred embodiments, A is selected from 4- to 7-memberednitrogen-containing heterocyclyl or 5- to 10-memberednitrogen-containing heteroaryl.

In some preferred embodiments, A is selected from 4- to 7-memberednitrogen-containing heterocyclyl or 5- to 10-memberednitrogen-containing heteroaryl, both of which are optionally substitutedwith a substituent selected from hydrogen, deuterium, C₁₋₆ alkyl andC₃₋₆ cycloalkyl that are optionally substituted with R⁶, halogen, —CN,halogenated C₁₋₆ alkyl, C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂heteroaryl; R⁶ is selected from deuterium, hydroxyl, —CN, C₁₋₆ alkoxyand C₃₋₆ cycloalkoxy.

In some preferred embodiments, A is selected from 5- or 6-memberednitrogen-containing heteroaryl.

In some preferred embodiments, A is selected from 5- or 6-memberednitrogen-containing heteroaryl optionally substituted with a substituentselected from deuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl that areoptionally substituted with R⁶, halogen, —CN, halogenated C₁₋₆ alkyl,C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl and C₅-12 heteroaryl; R⁶ is selectedfrom deuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆ cycloalkoxy.

In some preferred embodiments, A is a 5-membered nitrogen-containingheteroaryl.

In some preferred embodiments, A is a 5-membered nitrogen-containingheteroaryl optionally substituted with a substituent selected fromdeuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl that are optionallysubstituted with R⁶, halogen, —CN, halogenated C₁₋₆ alkyl, C₃₋₆heterocycloalkyl, C₆₋₁₀ aryl, and C₅₋₁₂ heteroaryl; R⁶ is selected fromdeuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆ cycloalkoxy;

preferably, A is a 5-membered nitrogen-containing heteroaryl substitutedwith a substituent, and the substituent is C₁₋₆ alkyl (e.g., propyl).

In some preferred embodiments, A is

wherein R⁴ is selected from hydrogen, deuterium, or C₁₋₆ alkyl and C₃₋₆cycloalkyl that are optionally substituted with R⁶, R⁶ is selected fromdeuterium, hydroxyl, —CN, C₁₋₄ alkoxy and C₃₋₆ cycloalkoxy, R⁵ isselected from hydrogen, deuterium, halogen, —CN, C₁₋₆ alkyl, halogenatedC₁₋₆ alkyl, C₁₋₆ alkylene-CN, C₁₋₆ alkylene-OH, C₁₋₆ alkylene-C₁₋₆alkoxy, C₃₋₆ cycloalkyl, C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂heteroaryl, X is selected from N and C—R⁵, each R⁵ is identical ordifferent.

In some preferred embodiments, A is selected from

In some preferred embodiments, R⁴ is C₁₋₆ alkyl optionally substitutedwith R⁶, R⁶ is selected from deuterium, hydroxyl, —CN, and C₁₋₄ alkoxy.

In some preferred embodiments, R⁴ is C₁₋₆ alkyl optionally substitutedwith R⁶, R⁶ is selected from hydroxyl and C₁₋₄ alkoxy.

In some preferred embodiments, R⁴ is C₁₋₄ alkyl optionally substitutedwith R⁶, R⁶ is selected from hydroxyl and C₁₋₄ alkoxy.

In some preferred embodiments, R⁴ is C₁₋₄ alkyl optionally substitutedwith R⁶, R⁶ is selected from hydrogen, hydroxyl, methoxy, and ethoxy.

In some preferred embodiments, R⁴ is C₁₋₄ alkyl optionally substitutedwith R⁶, R⁶ is selected from methoxy and ethoxy.

In some preferred embodiments, R⁴ is selected from n-propyl and

In some preferred embodiments, R⁵ is independently selected fromhydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl, C₁₋₆alkylene-OH, C₁₋₆ alkylene-C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₃₋₆heterocycloalkyl.

In some preferred embodiments, R⁵ is independently selected fromhydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl, and C₃₋₆cycloalkyl.

In some preferred embodiments, R⁵ is independently selected fromhydrogen, methyl, trifluoromethyl and cyclopropyl.

In some preferred embodiments, R⁵ is hydrogen.

In some preferred embodiments, X is selected from N and C—R⁵, R⁵ isselected from hydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl,C₁₋₆ alkylene-OH, C₁₋₆ alkylene-C₁₋₆ alkoxy, C₃₋₆ heterocycloalkyl, andC₃₋₆ heterocycloalkyl.

In some preferred embodiments, X is selected from N and C—R⁵, R⁵ isselected from hydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl,and C₃₋₆ cycloalkyl.

In some preferred embodiments, X is selected from N and C—R⁵, R⁵ isselected from hydrogen, methyl, trifluoromethyl, and cyclopropyl.

In some preferred embodiments, L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene and C₅₋₁₂ heteroarylene; preferably, L₁ isselected from C₁₋₃ alkylene and C₃₋₅ cycloalkylene; more preferably, L₁is selected from methylene and ethylene;

L₂ is absent or selected from —NH—, C₂₋₄ alkylene, C₂₋₃ alkenylene,—(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene,C₆₋₈ arylene, C₅₋₈ heteroarylene and 3- to 6-membered heterocyclylene;said —NH—, C₂₋₄ alkylene, C₂-3 alkenylene, —(C₁₋₃alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈arylene, C₅₋₈ heteroarylene, and 3- to 6-membered heterocyclylene areoptionally substituted with one or more substituents that areindependently selected from hydrogen, deuterium, halogen, hydroxyl,cyano, methyl, halomethyl, halomethoxy, —C(O)CH₃, —COORS and —NR³CO₂R³;said 3- to 6-membered heterocyclylene contains one or more ring membersselected from N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2; R³ is selected from hydrogen, deuterium, methyl and cyclopropyl;preferably, L₂ is absent or selected from —NH—, —N(CH₃)—, —C(CH₃)₂—,—CH═CH—, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene and 3- to 6-memberedheterocyclylene, wherein y₁ and y₂ are each independently selected from0, 1 or 2; more preferably, L₂ is absent or selected from —NH—, —N(CH₃)—and —C(CH₃)₂—;

L₃ is absent or selected from C₁₋₃ alkylene, C₂₋₃ alkenylene, C₂₋₃alkynylene, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene, and 3-to 7-membered heterocyclylene; preferably, L₃ is absent or selected fromC₁₋₆ alkylene and C₂₋₆ alkenylene; more preferably, L₃ is absent orselected from —C(CH₃)₂—, —CH₂—, and —CH═CH—; particularly preferably, L₃is absent or —CH₂—.

In some preferred embodiments, n is selected from 1 or 2, preferably, nis 1.

In some preferred embodiments, the present application provides acompound of general formula I_(a) or I_(b) or a pharmaceuticallyacceptable salt, ester, solvate (e.g., hydrate), stereoisomer, tautomer,prodrug thereof, or crystalline form, metabolite thereof, or a mixtureof the aforementioned:

wherein R¹, R⁴, R⁵, L₁, L₂, L₃, X and n are as defined above.

In some preferred embodiments, the present application providescompounds of formula II_(a) and formula II_(a′) or a pharmaceuticallyacceptable salt, ester, solvate (e.g., hydrate), stereoisomer, tautomer,prodrug thereof, or crystalline form, metabolite thereof, or a mixtureof the aforementioned:

wherein R¹, R⁴, R⁵, X, L₁, L₂, L₃ or n are as defined above.

In some preferred embodiments, the present application providescompounds of general formula II_(a) and formula II_(a′), wherein R¹ isselected from hydrogen, deuterium, and C₁₋₆ alkyl, said C₁₋₆ alkyl isoptionally substituted with R², R² is selected from hydrogen, deuterium,hydroxy, amino, carboxy, C₁₋₆ alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆alkyleneoxy)_(m2)-C₁₋₆ alkyl, (C₁₋₆ alkyl)₂ amino, and 5- to 6-memberedheterocyclyl containing one or more N, O or S atoms; wherein m₁ and m₂are each independently selected from 0, 1, 2, 3 or 4;

preferably, R¹ is C₁₋₆ alkyl, said C₁₋₆ alkyl is optionally substitutedby C₁₋₆ alkoxy;

preferably, R¹ is C₁₋₃ alkyl, said C₁₋₃ alkyl is optionally substitutedby C₁₋₆ alkoxy;

preferably, R¹ is ethyl, said ethyl is substituted with n-butoxy.

In some preferred embodiments, the present application providescompounds of general formula II_(a) and formula II_(a′), wherein X isselected from N and C—R⁵, R⁵ is selected from hydrogen, deuterium,halogen, —CN, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl, C₁₋₆ alkylene-CN, C₁₋₆alkylene-OH, C₁₋₆ alkylene-C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, C₃₋₆heterocycloalkyl, C₆₋₁₀ aryl, and C₅₋₁₂ heteroaryl;

preferably, X is selected from N and C—R⁵, R⁵ is selected from hydrogen,deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl and C₃₋₆ cycloalkyl;

preferably, X is selected from N and C—R⁵, R⁵ is selected from hydrogen,C₁₋₃ alkyl, halogenated C₁₋₃ alkyl and C₃₋₆ cycloalkyl;

preferably, X is selected from N and C—R⁵, R⁵ is selected from hydrogen,methyl, trifluoromethyl and cyclopropyl;

preferably, X is C—R⁵, R⁵ is selected from hydrogen and methyl.

In some preferred embodiments, the present application providescompounds of general formula II_(a) and formula II_(a′), wherein R⁴ isselected from hydrogen, deuterium, or C₁₋₆ alkyl and C₃₋₆ cycloalkyloptionally substituted with R⁶, R⁶ is selected from hydrogen, deuterium,hydroxyl, —CN, C₁₋₄ alkoxy and C₃₋₆ cycloalkoxy;

preferably, R⁴ is selected from C₁₋₆ alkyl and C₃₋₆ cycloalkyl, both ofwhich are optionally substituted with R⁶, R⁶ is C₁₋₄ alkoxy;

preferably, R⁴ is selected from C₁₋₃ alkyl optionally substituted withR⁶, R⁶ is C₁₋₃ alkoxy;

preferably, R⁴ is selected from propyl and methoxy-substituted ethyl;

preferably, R⁴ is n-propyl.

In some preferred embodiments, the present application providescompounds of general formulae II_(a) and II_(a), wherein:

L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene,C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene;

preferably, L₁ is C₁₋₃ alkylene; preferably, L₁ is selected frommethylene and ethylene.

In some preferred embodiments, the present application providescompounds of general formulae II_(a) and II_(a), wherein:

L₂ is absent or selected from —NH—, C₂₋₆ alkylene, C₂₋₆ alkenylene,—(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene,C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene and 3- to 10-memberedheterocyclylene; said —NH—, C₂₋₆ alkylene, C₂₋₆ alkenylene, —(C₁₋₆alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆-10arylene, C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene areoptionally substituted with one or more substituents that areindependently selected from hydrogen, deuterium, halogen, hydroxyl,cyano, C₁₋₃ alkyl, C₁₋₃ haloalkyl, C₁₋₃ haloalkoxy, —C(O)C₁₋₄ alkyl,—COOR³ and —NR³CO₂R³; said 3- to 10-membered heterocyclylene containsone or more ring members selected from N, NR³, O, P, and S(O)_(z);wherein y₁ and y₂ are each independently selected from 0, 1, 2, 3 or 4;z is selected from 0, 1 or 2; R³ is selected from hydrogen, deuterium,C₁₋₄ alkyl and C₃₋₆ cycloalkyl;

preferably, L₂ is absent or selected from —NH—, —N(CH₃)—, —C(CH₃)₂—,—CH═CH—, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene and 3- to 6-memberedheterocyclylene, wherein y₁ and y₂ are each independently selected from0, 1 or 2;

preferably, L₂ is absent or selected from —NH—, —N(CH₃)— and —C(CH₃)₂—;

preferably, L₂ is selected from —NH—, —N(CH₃)— and —C(CH₃)₂—.

In some preferred embodiments, L₂ is absent.

In some preferred embodiments, the present application providescompounds of general formulae II_(a) and II_(a′), wherein,

L₃ is absent or selected from C₁₋₆ alkylene, C₂₋₆ alkenylene, C₂₋₆alkynylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and3- to 10-membered heterocyclylene;

preferably, L₃ is absent or selected from C₁₋₃ alkylene, C₂₋₃alkenylene, C₂₋₃ alkynylene, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 7-membered heterocyclylene;

preferably, L₃ is absent or selected from C₁₋₆ alkylene and C₂₋₆alkenylene;

preferably, L₃ is absent or selected from —C(CH₃)₂—, —CH₂—, and —CH═CH—;

preferably, L₃ is absent.

In some preferred embodiments, the present application providescompounds of general formulae II_(a) and II_(a′), wherein,

n is selected from 0, 1 or 2; preferably, n is selected from 0 and 1;

preferably, n is 1.

In some preferred embodiments, the present application providescompounds of general formulae II_(a) and II_(a′), wherein:

R¹ is C₁₋₃ alkyl, said C₁₋₃ alkyl is optionally substituted with C₁₋₆alkoxy;

X is selected from N and C—R⁵, R⁵ is selected from hydrogen, methyl,trifluoromethyl and cyclopropyl;

R⁴ is selected from propyl and methoxy-substituted ethyl;

L₁ is C₁₋₃ alkylene; preferably, L₁ is selected from methylene andethylene;

L₂ is absent or selected from —NH—, —N(CH₃)— and —C(CH₃)₂—;

L₃ is absent or selected from —C(CH₃)₂—, —CH₂—, and —CH═CH—;

n is selected from 0 and 1; preferably, n is 1.

In some preferred embodiments, the present application providescompounds of general formulae II_(b) and formula II_(b′) or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned:

wherein R¹, R⁴, R⁵, X, L₁, L₂, L₃ or n are as defined above.

In some preferred embodiments, the present application provides acompound or a pharmaceutically acceptable salt, ester, solvate (e.g.,hydrate), stereoisomer, tautomer, prodrug thereof, or crystalline form,metabolite thereof, or a mixture of the aforementioned, wherein thecompound is selected from:

An atom in the compound of the invention may be replaced by itsisotopes. For example, ¹²C may be replaced by its isotope ¹³C or ¹⁴C; ¹Hmay be replaced by ²H (D, deuterium) or ³H (T, tritium), and so on. Theinvention includes an isotope-labeled compound obtained by replacing anyatom in the compound with its isotope.

The present application also relates to a process for the preparation ofa compound of formula I, comprising the following synthetic route:

wherein R¹, n, L₁, L₂, L₃ or A is as defined above, Lisa group bearingan aldehyde group at the end, and L may be converted to L₁ (preferably,L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selected from methyleneand ethylene) after reactions, and the carboxyl protection group PG isselected from the following groups that are optionally substituted withone or more R⁷: C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, C₃₋₁₀cycloalkyl, C₃₋₁₀ heterocycloalkyl, C₆₋₁₀ aryl, C₁₋₆ alkylene-C₆₋₁₀aryl, C₅₋₁₂ heteroaryl, R⁷ is selected from hydrogen, deuterium,halogen, C₁₋₆ alkyl and C₁₋₆ alkoxy. In some preferred embodiments, thecarboxyl protection group PG is preferably C₁₋₆ alkyl.

Particularly, the process comprises the following steps:

Step I: Reacting Compound a with Compound b to Provide Compound c:

the reaction is preferably carried out in a suitable organic solvent,and said organic solvent may be selected from a linear or cyclic ether(e.g. tetrahydrofuran or diethyl ether), a halogenated hydrocarbon (e.g.dichloromethane, chloroform, 1,2-dichloroethane or the like), a nitrile(e.g. acetonitrile or the like), N-methylpyrrolidone, dimethylformamide,dimethylacetamide, 1,4-dioxane, dimethylsulfoxide and any combinationthereof, preferably dioxane or 1,2-dichloroethane. The reaction ispreferably carried out in the presence of a suitable reducing agent, andsaid reducing agent is selected from sodium borohydride, potassiumborohydride, sodium cyanoborohydride and sodium borohydride acetate,preferably the reducing agent is sodium cyanoborohydride. The reactionis preferably carried out under suitable acidic condition, and said acidmay be selected from hydrochloric acid, acetic acid and trifluoroaceticacid, preferably the acid is trifluoroacetic acid. The reaction ispreferably carried out at a suitable temperature, and said temperatureis preferably from −50 to 100° C., more preferably from −20 to 70° C.The reaction is preferably carried out for a suitable time, for example1 to 24 hours, and for example 1 to 6 hours.

Step II: Reacting Compound c with Compound d to Provide Compound e.

The reaction is preferably carried out in a suitable organic solvent,and said organic solvent may be selected from a halogenated hydrocarbon(e.g., dichloromethane, chloroform, 1,2-dichloroethane or the like), anitrile (e.g., acetonitrile or the like), N-methylpyrrolidone,dimethylformamide, dimethylacetamide, 1,4-dioxane, dimethylsulfoxide,and any combination thereof, preferably dichloromethane. The reaction ispreferably carried out in the presence of a suitable condensing agent,and the condensing agent may be selected from thionyl chloride, oxalylchloride, phosphorus oxychloride, phosphorus trichloride, phosphoruspentachloride, ethyl chloroformate, isopropyl chloroformate, HATU, HBTU,EEDQ, DEPC, DCC, DIC, EDC, BOP, PyAOP or PyBOP, preferably thecondensing agent is thionyl chloride, oxalyl chloride. The reaction ispreferably carried out under suitable basic condition; the base is anorganic or inorganic base; preferably, the organic base is selected fromtriethylamine, DIPEA, pyridine, NMM or DMAP, and preferably, theinorganic base is selected from NaH, NaOH, Na₂CO₃ or K₂CO₃; preferably,the base is selected from triethylamine, DIPEA and DMAP. The reaction ispreferably carried out at a suitable temperature, and the reactiontemperature is preferably from 0 to 100° C., more preferably from 15 to50° C. The reaction is preferably carried out for a suitable time, forexample 1 to 24 hours, and for example 2 to 7 hours.

Step III: Removing the Protection Group of Compound e to Provide aCompound of General Formula I.

The reaction is preferably carried out in suitable water and/or anorganic solvent, and the organic solvent may be selected from ahalogenated hydrocarbon (e.g. dichloromethane, chloroform,1,2-dichloroethane or the like), a nitrile (e.g. acetonitrile or thelike), an alcohol (e.g. methanol, ethanol), an ether (e.g.tetrahydrofuran, dioxane), N-methylpyrrolidone, dimethylformamide,dimethylacetamide, dimethylsulfoxide and any combination thereof,preferably water and ethanol. The reaction is preferably carried outunder suitable acidic condition, and the acid is hydrochloric acid,sulfuric acid or trifluoroacetic acid. The reaction is preferablycarried out under suitable basic condition, and the base is sodiumhydroxide, potassium hydroxide, sodium carbonate, potassium carbonate orcesium carbonate, preferably the base is selected from sodium hydroxide.The reaction is preferably carried out at a suitable temperature, andthe reaction temperature is preferably from −20 to 100° C., morepreferably 0 to 60° C. The reaction is preferably carried out for asuitable time, for example 1 to 24 hours.

The application also relates to a process for the preparation of acompound of formula I_(a):

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃, PG and X are as defined above, L is agroup bearing an aldehyde group at the end, and L may be converted to L₁(preferably, L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selectedfrom methylene and ethylene), the process for the preparation of thecompound of formula I_(a) refers to the process for the preparation ofthe compound of formula I.

The application also relates to a process for the preparation of acompound of formula I_(b):

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃, PG and X are as defined above, L is agroup bearing an aldehyde group at the end, and L may be converted to L₁(preferably, L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selectedfrom methylene and ethylene), the process for the preparation of thecompound of formula I_(b) refers to the process for the preparation ofthe compound of formula I.

In some embodiments, the present application provides a process for thepreparation of the compounds of general formula II_(a) and/or generalformula II_(a′):

Process I:

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃, PG and X are as defined above, L is agroup bearing an aldehyde group at the end, and L may be converted to L₁(preferably, L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selectedfrom methylene and ethylene).

The synthetic steps of the process refer to the steps for the synthesisof the compound of formula I_(a).

Process II:

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃ and X are as defined above.

In particular, Process II comprises the following steps:

Step I: Separating the Compound of Formula I_(a) to Provide theCompounds of General Formula II_(a) and General Formula II_(a′).

The separation may be applied by using a commercially available chiralpreparative column, preferably OD, OJ, AS, AD, IA, IB, IC, ID, etc. withan alcohol (ethanol, isopropanol or the like), a weak polar solvent(n-hexane, petroleum ether, CO₂ or the like), a cosolvent(dichloromethane or the like) and a base (ammonia water, diethylamine orthe like) as the mobile phase to perform separations and purifications,and ingredients having ee value of >99% are collected.

Process III:

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃ and X are as defined above.

In particular, Process III comprises the following steps:

Step I: Separating Compound e to Provide Compound h and Compound h′:

The method is applied by using a commercially available chiralpreparative column, preferably OD, OJ, AS, AD, IA, IB, IC, ID, etc. withan alcohol (ethanol, isopropanol or the like), a weak polar solvent(n-hexane, petroleum ether, CO₂ or the like), a cosolvent(dichloromethane or the like) and a base (ammonia water, diethylamine orthe like) as the mobile phase to perform separations and purifications,and ingredients having ee value of >99% are collected.

Step II: Removing the Protection Groups of Compound h and Compound h′ toRespectively Provide Compounds of General Formula II_(a) and GeneralFormula II_(a′):

The reaction is preferably carried out in suitable water and/or anorganic solvent, and the organic solvent may be selected from ahalogenated hydrocarbon (e.g. dichloromethane, chloroform,1,2-dichloroethane or the like), a nitrile (e.g. acetonitrile or thelike), an alcohol (e.g. methanol, ethanol), an ether (e.g.tetrahydrofuran, dioxane), N-methylpyrrolidone, dimethylformamide,dimethylacetamide, dimethylsulfoxide and any combination thereof,preferably water and ethanol. The reaction is preferably carried outunder suitable acidic condition, and the acid is hydrochloric acid,sulfuric acid or trifluoroacetic acid. The reaction is preferablycarried out under suitable basic condition, and the base is sodiumhydroxide, potassium hydroxide, sodium carbonate, potassium carbonate orcesium carbonate, preferably the base is selected from sodium hydroxide.The reaction is preferably carried out at a suitable temperature, andthe reaction temperature is preferably from −20 to 100° C., morepreferably 0 to 60° C. The reaction is preferably carried out for asuitable time, for example 1 to 24 hours.

In some embodiments, the present application provides a process for thepreparation of a compound of general formula II_(b) and/or generalformula comprising the following steps:

Process I:

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃, PG and X are as defined above, L is agroup bearing an aldehyde group at the end, and L may be converted to L₁(preferably, L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selectedfrom methylene and ethylene).

The synthetic steps of the process refer to the steps for the synthesisof the compound of general formula Ib.

Process II:

wherein R¹, R⁴, R⁵, n, L₁, L₂, L₃ and X are as defined above.

In particular, Process II comprises the following steps:

Step I: separating the compound of formula Ib to provide the compoundsof the formula II_(b) and II_(b′),

The separation is applied by using a commercially available chiralpreparative column, preferably OD, OJ, AS, AD, IA, IB, IC, ID, etc. withan alcohol (ethanol isopropanol or the like), a weak polar solvent(n-hexane, petroleum ether, CO₂ or the like), a cosolvent(dichloromethane or the like) and a base (ammonia water, diethylamine orthe like) as the mobile phase to perform separations and purifications,and ingredients having ee value of >99% are collected.

In another aspect, the present application provides a pharmaceuticalcomposition, comprising the compound of the first aspect or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned, and one or morepharmaceutically acceptable carriers.

In some preferred embodiments, the pharmaceutical composition is in aform of tablets, capsules, lozenges, hard candies, powder, sprays,creams, ointments, suppositories, gels, pastes, lotions, ointments,aqueous suspensions, injectable solutions, elixirs and syrups.

In some preferred embodiments, the pharmaceutical composition furthercomprises one or more other drugs that prevent or treat a CCR2- and/orCCR5-mediated disease or condition, especially nonalcoholic fatty liverdisease (NAFLD) or the like.

In a further aspect, the present application provides a pharmaceuticalformulation, comprising the compound of the first aspect or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned, or said composition, andone or more pharmaceutically acceptable carriers.

In a further aspect, the present application provides a kit product,comprising:

a) a first container, comprising, as a first therapeutic agent, at leastone compound of the first aspect or a pharmaceutically acceptable salt,ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof,or crystalline form, metabolite thereof, or a mixture of theaforementioned, or, as a first pharmaceutical composition, thepharmaceutical composition;b) an optional second container, comprising, as a second therapeuticagent, at least one other therapeutic agent, or as a secondpharmaceutical composition, a pharmaceutical composition comprising saidother therapeutic agent; andc) an optional packaging specification.

In some preferred embodiments, the other therapeutic agent is selectedfrom drugs, other than the compounds described in this disclosure, thatare useful in the prevention or treatment of a CCR2- and/orCCR5-mediated disease or condition, especially nonalcoholic fatty liverdiseases (NAFLD) or the like.

In another aspect, the present application provides use of the compoundof the first aspect or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned, the pharmaceutical composition, the pharmaceuticalformulation or the kit product, in the manufacture of a medicament forthe prevention or treatment of a CCR2- and/or CCR5-mediated disease orcondition, especially nonalcoholic fatty liver diseases (NAFLD) or thelike.

In another aspect, the present application provides the compound of thefirst aspect or a pharmaceutically acceptable salt, ester, solvate(e.g., hydrate), stereoisomer, tautomer, prodrug thereof, or crystallineform, metabolite thereof, or a mixture of the aforementioned, thepharmaceutical composition, the pharmaceutical formulation or the kitproduct, for use in the prevention or treatment of a CCR2- and/orCCR5-mediated disease or condition, especially nonalcoholic fatty liverdisease (NAFLD) or the like.

In another aspect, the present application provides a method for theprevention or treatment of a CCR2- and/or CCR5-mediated disease orcondition, especially nonalcoholic fatty liver disease (NAFLD) or thelike, comprising administering to an individual in need thereof atherapeutically effective amount of the compound or a pharmaceuticallyacceptable salt, ester, solvate (e.g., hydrate), stereoisomer, tautomer,prodrug thereof, or crystalline form, metabolite thereof, or a mixtureof the aforementioned, the pharmaceutical composition, thepharmaceutical formulation or the kit product, and optionally furthercomprising administering to the individual in need thereof with anotherdrug that are used for the prevention or treatment of a CCR2- and/orCCR5-mediated disease or condition, especially nonalcoholic fatty liverdisease (NAFLD) or the like.

In the present application, a subject of the pharmaceutical compositionis to promote the administration to an organism, or to facilitate theabsorption of active ingredients thereby exerting biological activity.The carrier includes, but are not limited to, an ion exchanger, aluminumoxide, aluminum stearate, lecithin, a serum protein such as human serumalbumin, a buffer substance such as a phosphate, glycerol, sorbic acid,potassium sorbate, a mixture of partial glycerides of saturatedvegetable fatty acids, water, a salt or an electrolyte such as protaminesulfate, disodium hydrogen phosphate, potassium hydrogen phosphate,sodium chloride, a zinc salt, colloidal silica, magnesium trisilicate,polyvinylpyrrolidone, a cellulosic substance, polyethylene glycol,sodium carboxymethylcellulose, polyacrylates, beeswax, lanolin.

The present application provides that the compound according to thefirst aspect or a pharmaceutically acceptable salt, ester, solvate(e.g., hydrate), stereoisomer, tautomer, prodrug thereof, or crystallineform, metabolite thereof, or a mixture of the aforementioned,pharmaceutical compositions, pharmaceutical formulations or kits may beadministered by the following routes: parenteral, topical, intravenous,oral, subcutaneous, intraarterial, intradermal, transdermal, rectal,intracranial, intraperitoneal, intranasal, and intramuscular routes oras an inhalant. Optionally, the pharmaceutical composition may beadministered in combination with other reagents that have at least someeffect in the treatment of various diseases.

The present application provides the compound according to the firstaspect or a pharmaceutically acceptable salt, ester, solvate (e.g.,hydrate), stereoisomer, tautomer, prodrug thereof, or crystalline form,metabolite thereof, or a mixture of the aforementioned or thepharmaceutical composition may be formulated into various suitabledosage forms depending on the administration route.

The pharmaceutical composition or the suitable dosage form described inthe present application may contain from 0.01 mg to 1000 mg of thecompound.

For oral administration, the compound or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned or pharmaceutical compositions may be formulated into anyorally acceptable preparation forms, including, but not limited to,tablets, capsules, aqueous solutions or aqueous suspensions. Among them,carriers used in tablets generally include lactose and corn starch,additionally, lubricants such as magnesium stearate may also be added.Diluents used in capsules generally include lactose and dried cornstarch. For aqueous suspensions, active ingredients are usually used bymixing with suitable emulsifiers and suspending agents. Optionally,sweeteners, flavoring agents or coloring agents may be further addedinto the above oral preparations.

For skin tropical administration, the compound or a pharmaceuticallyacceptable salt, ester, solvate (e.g., hydrate), stereoisomer, tautomer,prodrug thereof, or crystalline form, metabolite thereof, or a mixtureof the aforementioned or the pharmaceutical composition may beformulated into suitable ointments, lotions or creams, wherein activeingredients are suspended or dissolved in one or more carriers. Thecarriers used in ointments include, but are not limited to, mineral oil,liquid petrolatum, white petrolatum, propylene glycol, polyethyleneoxide, polypropylene oxide, emulsifying wax and water; carriers used inlotions or creams include, but are not limited to, mineral oil, sorbitanmonostearate Tween 60, cetyl ester wax, hexadecene aromatic alcohol,2-octyldodecanol, benzyl alcohol and water.

The compound or a pharmaceutically acceptable salt, ester, solvate(e.g., hydrate), stereoisomer, tautomer, prodrug thereof, or crystallineform, metabolite thereof, or a mixture of the aforementioned or thepharmaceutical composition may also be administered in the form ofsterile injection preparations, including sterile injection water, oilysuspensions or sterile injection solutions. Among them, useful carriersand solvents include water, a Ringer's solution and an isotonic sodiumchloride solution. In addition, sterile non-volatile oils may also beused as a solvent or a suspending medium, such as mono-glycerides ordi-glycerides.

In the embodiments of the present application, suitable in vitro or invivo assays are performed to determine the efficacy of thepharmaceutical composition and if the administrations are suitable forthe treatment of an individual suffering from the disease or states.Examples of these assays are described in following non-limitingexamples in connection with specific diseases or medical treatments.Typically, an effective amount of the pharmaceutical compositionsufficient to achieve preventive or therapeutic effects is about 0.001mg/kg body weight/day to about 10,000 mg/kg body weight/day. Suitably,the dose is from about 0.01 mg/kg body weight/day to about 1000 mg/kgbody weight/day. The dose may be in a range of about 0.01 to 1000 mg/kgbody weight of a subject, more typically, 0.1 to 500 mg/kg body weightof a subject, every day, every two days or every three days. Exemplarytherapeutic regimen includes once administration every two days or onceadministration every week or once administration every month. Thepreparations are typically administered many times, and the interval ofa single dose may be one day, one week, one month or one year.Alternatively, the preparations may be administered in a form ofsustained release preparation, and in this case, a low administrationfrequency is required. The dose and the frequency may vary according tothe half-life of the preparations in the subject or the treatment isprophylactical or therapeutic. In prophylactical applications, arelatively low dose is administrated in a low interval frequency. Intherapeutic applications, sometimes, a relatively high dose have to beadministrated in a relatively short interval until the progress ofdiseases is delayed or halted, preferably until an individual exhibitspartial or full improvements in disease symptoms, and thereafter, apreventive regimen may be administrated to the patient.

Unless otherwise defined below, all the technical terms and scientificterms used herein are intended to have the same meaning as commonlyunderstood by a person skilled in the art. The mentioned technologiesherein are intended to refer those commonly understood in the art,including variations that are obvious for a person skilled in the artand equivalent substitutions. Even though the following terms arebelieved to be well understood by a person skilled in the art, they arestill defined herein for better explanation of the invention.

As used herein, the terms “include”, “comprise”, “have”, “contain”, or“relate” and other variations thereof herein, are inclusive or open anddo not exclude other not specified elements or method steps.

As used herein, the term “alkyl” is defined as a straight-chain orbranched aliphatic hydrocarbon group. In some embodiments, the alkyl has1 to 8, such as 1 to 4, carbon atoms. For example, as used herein, theterm “C₁₋₆ alkyl” refers to the straight-chain or branched group having1 to 6 carbon atoms (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl,isobutyl, sec-butyl, tert-butyl, n-pentyl, or n-hexyl), which isoptionally substituted with one or more (such as 1 to 3) suitablesubstituents such as halogen (referred to as “haloalkyl”, such as —CF₃,—C₂F₅, —CHF₂, —CH₂F, —CH₂CF₃, —CH₂C₁, —CH₂CH₂CF₃, etc.).

As used herein, the term “alkylene” is defined as a saturated divalenthydrocarbyl group derived from a straight-chain or branched saturatedhydrocarbyl group by removing two hydrogen atoms, and the alkylene groupcontains 1-10 carbon atoms, 1-6 carbon atoms, 1-4 carbon atoms, or 1-3carbon atoms, such as methylene (—CH₂—), ethylene (—CH₂CH₂—),isopropylene (—CH(CH₃)CH₂—) or the like, wherein the alkylene group maybe independently unsubstituted or substituted with one or moresubstituents described in the invention.

As used herein, the term “cycloalkyl” refers to a saturated orunsaturated, non-aromatic, monocyclic or polycyclic (such as bicyclic)hydrocarbon ring (e.g., monocycles, such as cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, orbicycles, including spiro, fused or bridged systems (such asbicyclo[1.1.1]pentyl, bicyclo[2.2.1]heptyl or the like), which isoptionally substituted with one or more (such as 1 to 3) suitablesubstituents. The cycloalkyl group has 3 to 15, for example, 3 to 10carbon atoms, 3 to 7 carbon atoms, or 3 to 6 carbon atoms. For example,as used herein, the term “C₃₋₇ cycloalkyl” refers to a saturated orunsaturated, non-aromatic, monocyclic or polycyclic (such as bicyclic)hydrocarbon ring having 3 to 7 ring-forming carbon atoms (e.g.,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl), which isoptionally substituted with one or more (such as 1 to 3) suitablesubstituents, for example methyl-substituted cyclopropyl.

As used herein, the term “C₃₋₁₀ cycloalkylene” is defined as a divalentC₃₋₁₀ alkyl residue, for example divalent methylene residue, such ascyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene,cycloheptylene or cyclooctylene, such as cyclopropylene, cyclobutylene,cyclopentylene, cyclohexylene, and particularly cyclopentylene,cyclohexylene.

As used herein, the term “alkoxy” refers to a linear, branched or cyclicsaturated monovalent hydrocarbon group having the formula of —O-alkyl,wherein the term “alkyl” is as defined above or it is a “cycloalkyl” asdefined below, e.g. C₁₋₆ alkyl, C₁₋₃ alkyl or C₃₋₆ cycloalkyl, such asmethoxy, ethoxy, n-propoxy, isopropoxy, cyclopropoxy, n-butoxy,isobutoxy, t-butoxy, sec-butoxy, cyclobutoxy, pentyloxy, isopentyloxy orn-hexyloxy, or their isomers.

As used herein, the term “heterocyclyl” refers to a saturated orunsaturated, monocyclic or bicyclic group which contains 2, 3, 4, 5, 6,7, 8 or 9 carbon atoms in the ring and one or more (e.g., one, two,three or four) heteroatom-containing groups selected from C(═O), 0, S,S(═O), S(═O)₂ and NR^(a), wherein R^(a) represents hydrogen atom, C₁₋₆alkyl or halogenated C₁₋₆ alkyl; the heterocycloalkyl may be connectedto the rest moiety of the molecule via any one of the carbon atoms orthe nitrogen atom (if any). For example, C₃₋₆ heterocyclyl, C₅₋₆heterocyclyl or C₃₋₇ heterocyclyl are the heterocyclyl having 3-6, 5-6or 3-7 carbon atoms. 3- to 10-membered heterocyclyl is the group having3 to 10 carbon atoms and heteroatoms in the ring, and 4- to 7-memberedheterocyclyl is the group having 4 to 7 carbon atoms and heteroatoms inthe ring, for example, but it is not limited to, oxiranyl, aziridinyl,azetidinyl, oxyethanyl, tetrahydrofuranyl, dioxolinyl, pyrrolidinyl,pyrrolidinonyl, imidazolidinyl, pyrazolinyl, pyrrolinyl,tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl,piperazinyl or trithianyl.

As used herein, the term “heteroarylene” refers to divalent heteroarylgroup, e.g., having 5-12, 5-10, or 5-8 carbon atoms.

As used herein, the term “aryl” refers to a full-carbon monocyclic orfused polycyclic aromatic group having a conjugated 7-electron system.For example, as used herein, the term “C₆₋₁₀ aryl” is intended to referto an aromatic group containing 6 to 10 carbon atoms, such as phenyl ornaphthyl. The aryl is optionally substituted with one or more (such as 1to 3) suitable substituents.

As used herein, the term “arylene” refers to an aryl group having twounsaturated valences, and having, e.g., 6-10 or 6-8 carbon atoms, forexample, phenylene.

As used herein, the term “heteroaryl” refers to a monocyclic, bicyclicor tricyclic aromatic ring system having 5, 6, 8, 9, 10, 11, 12, 13 or14 ring atoms, for example 5-10 ring atoms, and in particular 1 or 2 or3 or 4 or 5 or 6 or 9 or 10 carbon atoms, and comprising at least oneheteroatom which may be identical or different (said heteroatom, forexample, is oxygen, nitrogen or sulfur), additionally, in eachoccurrence, the group may be benzo-fused. In particular, the heteroarylgroup is selected from thienyl, furyl, pyrrolyl, oxazolyl, thiazolyl,imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl,thiadiazolyl and their benzo derivatives; or pyridyl, pyridazinyl,pyrimidinyl, pyrazinyl, triazinyl and their benzo derivatives.

As used herein, the term “halo” or “halogen” group is defined to includeF, Cl, Br or I.

As used herein, the term “alkenyl” refers to an aliphatic hydrocarbongroup containing at least one carbon-carbon double bond and one hydrogenatom replaced by a bond. The alkenyl can be a straight-chain orbranched, and it contains about 2 to about 15 carbon atoms. For example,the “C₂₋₆ alkenyl” herein contains about 2 to about 6 carbon atoms.Non-limiting examples of the alkenyl include ethenyl, propenyl,n-butenyl, 3-methyl-but-2-enyl, n-pentenyl, octenyl and decenyl. Thealkenyl may be unsubstituted or substituted with one or moresubstituents that may be identical or different, each substituent isindependently selected from halogen, alkenyl, alkynyl, aryl, cycloalkyl,cyano, hydroxy, —O-alkyl, —O-aryl, -alkylene-O-alkyl, alkylthio, —NH₂,—NH(alkyl), —N(alkyl)₂, —NH(cycloalkyl), —O—C(O)-alkyl, —O—C(O)-aryl,—O—C(O)-cycloalkyl, —C(O)OH and —C(O)O-alkyl. The term “C₂₋₆ alkenyl”refers to an alkenyl group of 2 to 6 carbon atoms.

As used herein, the term “alkenylene” refers to a straight-chain orbranched divalent hydrocarbon group for connecting the rest moiety ofthe molecule, which only consists of carbon and hydrogen atoms andcontains at least one double bond and two to six carbon atoms, such asethenylene, propenylene, n-butenylene or the like. The alkenylene chainis connected to the rest moiety of the molecule via a double or singlebond and connected to a group via a double or single bond. Theconnection site of the alkenylene chain to the rest moiety of themolecular and the group may pass through one carbon atom or any twocarbon atoms in the chain. Unless otherwise specified in thedescription, the alkenylene chain may be optionally substituted with oneor more substituents independently selected from halo, cyano, nitro,aryl, cycloalkyl, heterocyclyl, heteroaryl, oxo, trimethylsilyl, —OR⁸,—OC(═O)R⁸, —N(R⁸)₂, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)N(R⁸)₂, —N(R⁸)C(═O)OR⁸,—N(R⁸)C(═O)R⁸, —N(R⁸)S(═O)_(t)R⁸ (where t is 1 or 2), —S(═O)_(t)OR⁸(where t is 1 or 2), —S(═O)_(p)R⁸ (where p is 0, 1 or 2), and—S(═O)_(t)N(R⁸)₂ (where t is 1 or 2), wherein each R⁸ is independentlyhydrogen, alkyl, haloalkyl, cycloalkyl, cycloalkylalky, aryl, arylalkyl,heterocyclyl, heterocyclylalkyl, heteroaryl, or heteroarylalkyl, andwherein each of the above substituents, as defined herein, may beoptionally substituted.

As used herein, the term “alkynyl” refers to a hydrocarbon group havingone or more C═C triple bonds. The alkynyl has, but is not limited to,2-18 carbon atoms, for example 2-10 carbon atoms, and for example 2-6carbon atoms. The “lower alkynyl” herein refers to an alkynyl havingless carbon atoms, for example, 2-8 carbon atoms, for example 2-6 carbonatoms, and for example 2-4 carbon atoms. Examples of the alkynyl hereininclude, but are not limited to, ethynyl, 2-propynyl, 2-butynyl, and1,3-butynyl. When the alkynyl as defined herein involves a numericalrange, for example “C₂-C₆ alkynyl” or “C₂₋₆ alkynyl”, it refers to analkynyl containing 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5carbon atoms, or 6 carbon atoms, and the alkynyl herein also encompassesinstances, numerical ranges of which are not specified.

As used herein, the term “alkynylene” refers to the alkynyl as definedabove in which one hydrogen atom has been replaced by a bond.Non-limiting examples of the alkynylene include —CH₂C≡C—, —CH₂C≡CCH₂—and —CH(CH₃)C≡C—, and for example, the alkynylene has 2 to 6 carbonatoms. The alkynylene is branched or straight. The term “C₂₋₆alkynylene” refers to an alkynylene group having 2 to 6 carbon atoms,and the term “C₂₋₃ alkynylene” refers to an alkynylene group having 2 to3 carbon atoms.

As used herein, the term “amino” refers to a nitrogen-containing moietyhaving two further substituents, wherein a chlorine or carbon atom isconnected to the nitrogen. For example, representative amino groupsinclude —NH₂, —NHCH₃, —N(CH₃)₂, —NHC₁₋₃ alkyl, —N(C₁₋₃ alkyl)₂ or thelike. Unless otherwise specified, a compound of the invention containingan amino moiety may include a derivative thereof in which the aminomoiety is protected. Suitable protecting groups of the amino moietyinclude acetyl, t-butyloxycarbonyl, benzyloxycarbonyl or the like.

As used herein, the term “carboxylic acid group,” also known as“carboxyl” refers to a functional group having the chemical formula of—COOH. In the sense of the present invention, the term “carboxyl”further includes the related anion —COO— (carboxylate ion). Depending onenvironmental conditions, for example pH value of the environment, thefunctional group may be present in the form of —COOH or —COO—.

As used herein, the term “halogenated C₁₋₆ alkyl” refers to a groupobtained from the C₁₋₆ alkyl as described above by substituting one ormore (e.g., 1, 2, 3 or 4) hydrogen atoms with halogen (e.g., fluorine,chlorine, bromine, or iodine), for example, halogenated C₁₋₃ alkyl,halomethyl, haloethyl, halogenated C₃ alkyl, halogenated C₄ alkyl,fluoro C₁₋₃ alkyl, fluoromethyl, fluoroethyl, fluoro C₃ alkyl, fluoro C₄alkyl, chloro C₁₋₃ alkyl, chloromethyl, chloroethyl, chloro C₃ alkyl,and chloro C₄ alkyl. Specific examples include, but are not limited to,fluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl,chloroethyl or the like.

The term “substituted” refers to that one or more (e.g., one, two,three, or four) hydrogen atoms on an indicated atom are selectivelyreplaced by indicated groups, provided that the normal valence of theindicated atom in current situations is not exceeded and thesubstitution can result in a stable compound. A combination ofsubstituents and/or variables is permissible only when such acombination can result in a stable compound.

The term “optionally substituted” refers to an optional substitutionwith a specified group, radical or moiety.

In the case that the bond of a substituent is shown as passing through abond connecting two atoms in a ring, such a substituent can be bonded toany ring-forming atom in the substitutable ring.

The compound of the invention may also comprise one or more (e.g., one,two, three, or four) isotopic replacements. For example, in saidcompound, H may be in any isotopic forms, including ¹H, ²H (D ordeuterium), and ³H (T or tritium); C may be in any isotopic forms,including ¹²C, ¹³C, and ¹⁴C; 0 may be in any isotopic forms, including¹⁶O and ¹⁸O.

The term “stereoisomer” refers to an isomer formed as a result of atleast one asymmetric center. In a compound having one or more (e.g.,one, two, three, or four) asymmetric centers, a racemate, a racemicmixture, a single enantiomer, a diastereomeric mixture and a singlediastereomer may be formed. A specified individual molecule may alsoexist in the form of a geometric isomer (cis/trans). Similarly, thecompound of the invention may be present as a mixture of two or moredifferent structural forms in a rapid equilibrium (commonly referred toas tautomers). Representative examples of tautomers include ketone-enoltautomers, phenol-ketone tautomers, nitroso-oxime tautomers,imine-enamine tautomers or the like. It shall be understood that thescope of the present application encompasses all such isomers ormixtures in any ratios (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%,96%, 97%, 98%, 99%).

The present application encompasses all possible crystalline forms orpolymorphs of the compound, and it may be a single polymorph polymorphor a mixture of more than one polymorphs in any ratios.

The compound or a pharmaceutically acceptable salt thereof may also forma solvate, such as an alcoholate or the like.

The compound may also be in the form of a prodrug or a form which wouldrelease the active ingredient after metabolism in vivo, may release theactive ingredient. Selecting and preparing a suitable prodrug derivativeis well-known techniques for a person skilled in the art.

The compound may also be in a chemically protected form. A protectinggroup can protect an active group (e.g. an amino group) of the compound,and the protecting group may be metabolized in vivo thus to releasecorresponding active ingredient. Selecting and preparing suitablechemically protected forms are well-known techniques for a personskilled in the art.

The term “pharmaceutically acceptable” as used in the presentapplication refers to that a substance or composition has to bechemically and/or toxicologically compatible with other components thatform a preparation and/or mammals to be treated with.

The term “pharmaceutically acceptable salt” as used in the presentapplication includes a conventional salt formed with a pharmaceuticallyacceptable inorganic or organic acid, or inorganic or organic base.Illustrative examples of suitable salts include, but are not limited to,organic salts derived from amino acids such as glycine and arginine,ammonia, primary, secondary and tertiary amines, and cyclic amines suchas piperidine, morpholine, and piperazine, and inorganic salts derivedfrom sodium, calcium, potassium, magnesium, manganese, iron, copper,zinc, aluminum, and lithium.

It also should be understood that some compounds of the presentapplication may be present in a free form or, when appropriate, in theform of a pharmaceutically acceptable derivative thereof. Thepharmaceutically acceptable derivative includes, but is not limited to,a pharmaceutically acceptable salt, a solvate, a metabolite, or aprodrug, and all of which can directly or indirectly provide thecompound, or a metabolite or residue thereof when being administered toa patient in need thereof.

The pharmaceutically acceptable salt of the compound includes an acidaddition salt and a base addition salt. Suitable acid addition salts areformed from acids capable of forming non-toxic salts.

Suitable base addition salts are formed from bases capable of formingnon-toxic salts. Acceptable inorganic bases include aluminum hydroxideand calcium hydroxide. Acceptable organic bases include tromethamine,N-methylglucamine or the like.

For reviews of suitable salts, please refer to Stahl and Wermuth,Handbook of Pharmaceutical Salts: Properties, Selection, and Use(Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptablesalts of the compounds are known for a person skilled in the art.

The compound may be present in a form of a hydrate or a solvate, whereinthe compound of the invention comprises, as a structural element of thecrystal lattice of the compound, a polar solvent, in particular, such aswater, methanol or ethanol. The amount of the polar solvent,particularly water, may be present in a stoichiometric ornon-stoichiometric ratio.

Also included within the scope of the present invention is a metaboliteof the compound of the invention, i.e., a compound formed in vivo upondrug administration.

The term “pharmaceutical composition” as used in the present applicationincludes a product comprising a therapeutically effective amount of thecompound, as well as any product formed, directly or indirectly, from acombination comprising the compound.

The term “effective amount” as used in the present application refers toan amount sufficient to achieve a desired therapeutic effect, e.g., anamount capable of achieving alleviation of symptoms associated withdiseases to be treated.

The term “treatment” as used in the present application aims toalleviate or eliminate a state or condition of the disease in interest.If one or more indications and symptoms of a subject are decreased orimproved observably and/or detectably after the subject receives atherapeutic amount of a compound, an optical isomer or apharmaceutically acceptable salt thereof, or a pharmaceuticalcomposition thereof according to the method as described herein, thesubject is successfully “treated”. It should be also understood that thetreatment of a disease state or condition not only includes completetreatment, but also includes the situation where the complete treatmentis not achieved, but some related biological or medical results areachieved.

“Treatment” refers to any administration of the compound, thus toachieve:

(1) preventing the occurrence of a disease in an animal that may bepredisposed to the disease but has not yet experienced or exhibitedpathology or symptomology of the disease;(2) inhibiting a disease (i.e., arresting further development ofpathology and/or symptomatology) in an animal that is experiencing orexhibiting the pathology or symptomatology of the disease; or(3) improving a disease (i.e., reversing pathology and/orsymptomatology) in an animal that is experiencing or exhibiting thepathology or symptomatology of the disease.

Technical Effects

Through deep researches, a nitrogen-containing benzoheterocycle compoundcontaining a carboxylic acid group is obtained. Via the introduction ofthe carboxylic acid group, the compound can achieve at least one of thefollowing technical effects:

(1) extremely remarkable liver targeting properties;(2) excellent pharmacokinetic properties;(3) potent inhibitory effect against CCR2/CCR5; and(4) remarkable treating effect for a CCR2- and/or CCR5-mediated relevantdisease, especially nonalcoholic fatty liver disease (NAFLD) or thelike.

SPECIFIC MODES FOR CARRYING OUT THE INVENTION

Embodiments of the present invention will be described in detail belowwith reference to examples. However, it will be understood by a personskilled in the art that the following examples are only used forillustration of the invention, but should be deemed as limitation to thescope of the invention. The examples, not indicated with specificconditions, should be performed according to conventional conditions orconditions recommended by manufacturers. The reagents or instruments,not indicated with manufacturers, each are conventional commerciallyavailable products.

In conventional synthesis and examples, and synthesis examples ofintermediates, the meanings of abbreviations are shown as follows:

Abbreviations Meaning Abbreviations Meaning OD Polysaccharide EEDQ2-Ethoxy-1-ethoxycarbonyl-1,2- derivative normal dihydroquinoline phasecoating type chiral chromatographic column, with a silica gel surfacecoated with cellulose-tri (3,5-dimethylphenyl carbamate) OJPolysaccharide DEPC Diethyl derivative normal cyanophosphonate phasecoating type chiral chromatographic column, with a silica gel surfacecoated with cellulose- tri(4-methyl benzoate) AS Polysaccharide DCCDicyclohexylcarbodiimide derivative normal phase coating type chiralchromatographic column, with a silica gel surface coated withamylose-tri(S)- α-methylphenyl carbamate AD Polysaccharide DIC N,N'-derivative normal Diisopropylcarbodiimide phase coating type chiralchromatographic column, with a silica gel surface coated withamylose-tri (3,5-dimethylphenyl carbamate) IA Solvent-resistant EDC1-(3- chiral column, with a Dimethylaminopropyl)-3- silica gel surfaceethylcarbodiimide covalently bonded to amylose-tris (3,5-dimethylphenylcarbamate) IB (IB) Solvent-resistant BOP Benzotriazole-1-tris chiralcolumn, with a (trimethylamino)-trifluorophosphate silica gel surfacecovalently bonded to cellulose-tri (3,5-dimethyl phenyl carbamate) ICSolvent-resistant PyAOP (3H-1,2,3-triazolo[4,5-B] chiral column, with apyridine-3-oxy)tri-1- silica gel surface pyrrolidyl covalently bonded tohexafluorophosphate cellulose-tri (3,5-dichlorophenyl carbamate) IDSolvent-resistant PyBOP 1H-benzotriazol-1- chiral column, with ayloxytripyrrolidinyl silica gel surface hexafluorophosphate covalentlybonded to mylose-tri (3-chlorophenyl carbamate) ee Enantiomeric excessDIPEA N,N-Diisopropylethylamine percentage HATUO-(7-azabenzotriazol-1-yl)- NMM N-Methylmorpholine N,N,N′,N′-tetramethyluronium hexafluorophosphate HBTU Benzotriazole-N,N,N′,N′-DMAP p-Dimethylaminopyridine tetramethyluronium hexafluorophosphate

Examples and Assays are exemplified below to further illustrate theinvention in detail, but not intend to limit the invention. In addition,variations may be made without departing from the scope of the presentinvention.

The structures of the compounds as described in the following exampleswere confirmed by a nuclear magnetic resonance (¹H NMR) or a massspectrometry (MS).

The measuring instrument of the nuclear magnetic resonance (¹H NMR) wasa Bruker 400 MHz nuclear magnetic resonance meter; the measuringsolvents were deuterated methanol (CD₃OD), deuterated chloroform (CDCl₃)and hexadeuterated dimethyl sulfoxide (DMSO-d₆); the internal standardsubstance was tetramethylsilane (TMS).

The abbreviations in the nuclear magnetic resonance (NMR) spectra usedin the examples are shown below:

s: singlet, d: doublet, t: triplet, q: quartet, dd: doublet double, qd:quartet doublet, ddd: double double double, ddt: double double triplet,dddd: double double double doublet, m: multiplet, br: broad, J: couplingconstant, Hz: hertz, DMSO-d₆: deuterated dimethyl sulfoxide.

All δ values are expressed in ppm.

The measuring instrument of the mass spectrometry (MS) was an Agilent(ESI) mass spectrometer, with the model Agilent 6120B.

EXAMPLES Example 1: Synthesis of(S,E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid (Compound 1)

Step I: Synthesis of ethyl 3-hydroxy-2,2-dimethylpropionate

At room temperature, compound 1-1 (1.0 g, 8.5 mmol), cesium carbonate(3.3 g, 10.2 mmol), and N,N-dimethylformamide (15 mL) were charged intoa three-necked flask, and next, iodoethane (2.6 g, 17.0 mmol) was addedthereto under stirring at room temperature. The system was replaced withnitrogen gas three times and heated to 55° C., and reacted for 16 hoursat the temperature. After being cooled to room temperature, the systemwas poured into ice water (80 mL) and extracted with ethyl acetate (30mL×3). The organic phase was washed with saturated brine (30 mL×3) anddried over anhydrous sodium sulfate, and the drying agent was removed byfiltration. The solvents were removed by evaporation at reducedpressure, and purification with a column chromatography (THF:PE=1:10)was performed to give 1.0 g of the title compound. ESI-MS (m/z): 147.1[M+H]⁺.

Step II: Synthesis of ethyl 2,2-dimethyl-3-oxopropionate

Oxalyl chloride (522 mg, 4.1 mmol) was dissolved in dichloromethane (5mL), and the system was cooled to −78° C. Dimethyl sulfoxide (640 mg,8.2 mmol) was dropwise added thereto. And after the addition, the systemwas reacted for 30 min at the temperature. Then, a dichloromethane (5mL) solution of compound 1-2 (500 mg, 3.4 mmol) was dropwise added. Andafter the addition, the system was reacted for 2 h at −78° C. Further,triethylamine (1.03 g, 10.2 mmol) was added dropwise. And after theaddition, the system was reacted for 1 h at −78° C. and then a further30 min at room temperature. The reaction was quenched by adding water(30 mL) and extracted with dichloromethane (20 mL×2). The organic phasewas washed with saturated brine (30 mL×2) and dried over anhydroussodium sulfate, and the drying agent was removed by filtration. Solventswere removed by evaporation under reduced pressure, and purificationwith a column chromatography (THF:PE=1:15) was performed to give 200 mgof the title compound. ESI-MS (M/z) 145.1 [M+H]⁺.

Step III: Synthesis of(E)-8-(4-(2-butoxyethoxy)phenyl)-1-(3-ethoxy-2,2-dimethyl-3-oxopropyl)-1,2,3,4-tetrahydrobenzo[b]azacyclooctatetraene-5-formicacid

At room temperature,(E)-8-(4-(2-butoxyethoxy)phenyl)-1,2,3,4-tetrahydrobenzo[b]azacyclooctatetraene-5-formicacid (80 mg, 0.2 mmol) and compound 1-3 (432 mg, 3.0 mmol) weredissolved in dioxane (4.5 mL), then trifluoroacetic acid (1.5 mL) wasadded thereto at room temperature, and after 5 min stirring, sodiumcyanoborohydride (252 mg, 4.0 mmol) was further added. After theaddition, the mixture was reacted at 60° C. for 2.5 h under microwave.The system was added with water (30 mL) and extracted with ethyl acetate(30 mL×2). The organic phase was washed with saturated brine (30 mL×2)and dried over anhydrous sodium sulfate, and the drying agent wasremoved by filtration. Solvents were removed by evaporation underreduced pressure, and preparative liquid phase purification wasperformed to give 150 mg of the title compound. ESI-MS (M/z) 524.3[M+H]⁺.

Step IV: Synthesis of ethyl (5,E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropionate

Compound 1-4 (60 mg, 0.11 mmol) was dissolved in dichloromethane (2 mL)and added with thionyl chloride (0.5 mL) at room temperature, and themixture was reacted for 1 h. The system was drained, and then dissolvedby adding dichloromethane (2 mL), the resulting mixture was dropwiseadded to a dichloromethane (2 mL) solution of(S)-4-(((1-propyl-1H-imidazol-5-yl)methyl)sulfinyl) aniline (46 mg, 0.16mmol) and 4-dimethylaminopyridine (84 mg, 0.66 mmol). After theaddition, the mixture was reacted at room temperature for 3 h. Thereaction was quenched by adding water (10 mL) and extracted withdichloromethane (20 mL×2). The organic phase was washed with saturatedbrine (20 mL×2) and dried over anhydrous sodium sulfate, and the dryingagent was removed by filtration. Solvents were removed by evaporationunder reduced pressure, and purification was performed on preparativehigh performance liquid chromatography to give 15 mg of the titlecompound. ESI-MS (M/z) 769.4 [M+H]⁺.

Step V: Synthesis of(S,E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid

At room temperature, compound 1-5 (15 mg, 0.02 mmol) was dissolved inanhydrous ethanol (0.5 mL) and water (1 mL), and sodium hydroxide (8 mg,0.2 mmol) was added thereto under stirring at room temperature. Themixture was warmed to 45° C. and reacted for 20 h at the temperature.The reaction was detected by LC-MS to show the completion of thereaction. After cooling to room temperature, the system was added with1N hydrochloric acid aqueous solution to adjust the pH to 5-6 and thenextracted with ethyl acetate (20 mL×2). The organic phase was washedwith saturated brine (20 mL×1) and dried over anhydrous sodium sulfate,and the drying agent was removed by filtration. The solvents wereevaporated under reduced pressure, and purification was performed onpreparative high performance liquid chromatography to give 5 mg of thetitle compound 1.

The structure of compound 1 was characterized as follows:

¹H NMR (400 MHz, DMSO-d₆) δ 10.07 (s, 1H), 7.89 (d, J=8.5 Hz, 2H), 7.63(s, 1H), 7.55 (d, J=8.6 Hz, 2H), 7.51 (d, J=2.4 Hz, 1H), 7.47 (d, J=9.2Hz, 4H), 7.12 (d, J=8.8 Hz, 1H), 7.00 (d, J=8.5 Hz, 2H), 6.48 (s, 1H),4.35 (d, J=14.3 Hz, 1H), 4.17 (d, J=14.3 Hz, 1H), 4.11 (dd, J=5.8, 3.6Hz, 2H), 3.80 (td, J=7.1, 2.2 Hz, 2H), 3.74-3.68 (M, 2H), 3.49-3.42 (M,6H), 2.38 (d, J=6.2 Hz, 2H), 1.65 (q, J=7.3 Hz, 2H), 1.55-1.45 (M, 4H),1.39-1.27 (M, 2H), 1.17 (s, 6H), 0.88 (t, J=7.3 Hz, 3H), 0.81 (t, J=7.4Hz, 3H). ESI-MS (M/z): 741.4 [M+H]⁺.

Example 2: Synthesis of(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((4-methyl-1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid (Compound 2)

Step I: Synthesis of 4-methyl-1-propyl-1H-imidazol-5-yl-formaldhyde

At room temperature, compound 2-1 (500 mg, 4.5 mmol), potassiumcarbonate (941 mg, 6.8 mmol) and N,N-dimethylformamide (10 mL) wereadded into a three-necked flask, and iodopropane (1.5 g, 9.1 mmol) wasadded thereto. Under the protection of nitrogen gas, the mixture waswarmed to 50° C. and reacted for 16 h at the temperature. The system wascooled to room temperature, poured into ice water (100 mL) and extractedwith ethyl acetate (40 mL×3). The organic phase was washed withsaturated saline (30 mL×3), dried over anhydrous sodium sulfate, andconcentrated, and purified by column chromatography to give 260 mg ofthe title compound. ESI-MS (M/z) 153.1 [M+H]⁺.

Step II: Synthesis of (4-methyl-1-propyl-1H-imidazol-5-yl) methanol

Compound 2-2 (210 mg, 1.4 mmol) was dissolved in anhydrous methanol (5mL) and cooled to 0° C., and sodium borohydride (66 mg, 1.7 mmol) wasadded thereto in batches. The mixture was reacted at the temperature for30 min. The system was added with water (10 mL) to quench the reactionand concentrated, and ethyl acetate (30 mL×2) was added thereto forextraction. The organic phase was washed with saturated brine (10 mL×2),dried over anhydrous sodium sulfate, and concentrated to give 155 mg ofthe title compound. ESI-MS (M/z): 155.1 [M+H]⁺.

Step III: Synthesis of 5-(chloromethyl)-4-methyl-1-propyl-1H-imidazole

Compound 2-3 (155 mg, 1.0 mmol) was dissolved in acetonitrile (3 mL),and thionyl chloride (1 mL) was added thereto at room temperature. Then,the mixture was warmed to 70° C. and reacted for 2 h. The system wascooled to room temperature, concentrated, added with ethyl acetate (5mL), and stirred at room temperature for 30 minutes. Suction filtrationwas performed to give 175 mg of the hydrochloride of the title compound.¹H NMR (400 MHz, DMSO-d₆) δ 14.93 (s, 1H), 9.14 (s, 1H), 5.06 (s, 2H),4.15 (t, J=7.4 Hz, 2H), 2.34 (s, 3H), 1.86 (H, J=7.4 Hz, 2H), 0.91 (t,J=7.4 Hz, 3H).

Step IV: Synthesis of4-(((4-methyl-1-propyl-1H-imidazol-5-yl)methyl)thio) aniline

At room temperature, 4-aminothiophenol (240 mg, 1.9 mmol), triethylamine(576 mg, 5.7 mmol) and tetrahydrofuran (10 mL) were added to athree-necked flask and cooled to 0° C., and a tetrahydrofuran (5 mL: 0.5mL) aqueous solution of compound 2-4 (400 mg, 1.9 mmol) was addeddropwise thereto. After the addition, the mixture was warmed to roomtemperature and reacted for 2 hours. The system was poured into water(50 mL) and extracted with ethyl acetate (40 mL×3). The organic phasewas washed with saturated brine (30 mL×3), dried over anhydrous sodiumsulfate, and concentrated. Purification was performed on a preparativehigh performance liquid chromatography to give 160 mg of the titlecompound. ESI-MS (M/z) 262.1[M+H]⁺.

Step V: Synthesis of4-(((4-methyl-1-propyl-1H-imidazol-5-yl)methyl)sulfinyl) aniline

Compound 2-5 (160 mg, 0.61 mmol) was dissolved in dichloromethane (5 mL)and cooled to 0° C., and m-chloroperoxybenzoic acid (105 mg, 0.61 mmol)was added thereto in batches. After the addition, the mixture wasreacted for 30 min at the temperature. The system was added withsaturated anhydrous sodium sulfite solution (10 mL) to quench thereaction, and extracted by adding dichloromethane (20 mL×3). The organicphase was washed with saturated brine (20 mL×2), dried over anhydroussodium sulfate, and concentrated. Purification was performed onpreparative high performance liquid chromatography to give 180 mg of thetitle compound. ESI-MS (M/z) 278.1 [M+H]⁺.

Step VI: Synthesis of methyl(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((4-methyl-1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropionate

The synthesis was performed by applying a method similar to Step IV inExample 1, except that(S)-4-(((1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)aniline was replacedwith compound 2-6, and purification was performed on columnchromatography to give 230 mg of the title compound. ESI-MS (M/z) 769.4[M+H]⁺

Step VII: Synthesis of(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((4-methyl-1-propyl-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid

The synthesis was performed by applying a method similar to Step V inExample 1, except that compound 1-5 was replaced with compound 2-7, andpurification was performed on preparative high performance liquidchromatography to give 60 mg of the title compound 2.

The structure of compound 2 was characterized as follows:

¹H NMR (400 MHz, DMSO-d₆) δ 12.45 (s, 1H), 10.08 (s, 1H), 7.87 (d, J=8.5Hz, 2H), 7.53 (t, J=9.2 Hz, 4H), 7.50-7.44 (m, 2H), 7.39 (d, J=8.5 Hz,2H), 7.12 (d, J=8.9 Hz, 1H), 7.00 (d, J=8.6 Hz, 2H), 4.26 (d, J=14.3 Hz,1H), 4.16 (d, J=14.3 Hz, 1H), 4.13-4.07 (m, 2H), 3.79 (ddd, J=18.6,13.9, 6.8 Hz, 2H), 3.73-3.66 (m, 2H), 3.52-3.40 (m, 4H) 3.29 (s, 2H),2.38 (s, 2H), 1.70-1.63 (M, 2H), 1.61 (s, 3H), 1.50 (dt, J=13.8, 6.2 Hz,4H), 1.36-1.29 (M, 2H), 1.17 (s, 6H), 0.88 (t, J=7.3 Hz, 3H), 0.82 (t,J=7.1 Hz, 3H). ESI-MS (M/z): 755.3 [M+H]⁺.

Example 3

Compound 2 (260 mg) obtained in Example 2 was separated on chiral columnfor one of its enantiomers by applying SFC method, and the correspondingfraction was collected. The solvent was removed by rotary evaporation togive the pure optical isomer. The preparation conditions are as follows:chiral column: CHIRALPAK ID; chiral column model: 0.46 cm I.D.×15 cm L;sampling quantity: 0.5 uL; mobilephase:CO₂/(EtOH:DCM:NH₄OH=60:40:0.1)=55/45 (V/V); flow rate: 2.0 ml/min;wavelength: UV 254 nm; 100 mg of the isomer A of compound 2 was obtainedwith a retention time of 4.773 min.

The structure of the isomer A was characterized as follows:

¹H NMR (400 MHz, DMSO-d₆) δ 12.45 (s, 1H), 10.09 (s, 1H), 7.87 (d, J=8.7Hz, 2H), 7.58-7.50 (m, 4H), 7.49 (s, 1H), 7.48-7.44 (m, 1H), 7.39 (d,J=8.7 Hz, 2H), 7.12 (d, J=8.8 Hz, 1H), 7.02-6.97 (m, 2H), 4.26 (d,J=14.4 Hz, 1H), 4.16 (d, J=14.4 Hz, 1H), 4.13-4.08 (m, 2H), 3.84-3.75(m, 2H), 3.70 (dd, J=5.6, 3.7 Hz, 2H), 3.50-3.42 (m, 4H), 3.29 (s, 2H),2.38 (s, 2H), 1.70-1.63 (M, 2H), 1.61 (s, 3H), 1.54-1.45 (M, 4H),1.36-1.30 (M, 2H), 1.17 (s, 6H), 0.88 (t, J=7.4 Hz, 3H), 0.82 (t, J=7.3Hz, 3H). ESI-MS (M/z): 755.3 [M+H]⁺.

specific optical rotation: [α]_(280nm) ^(20° C.)=+58.0 (c=0.044, MeOH).

Example 4

Compound 2 (260 mg) obtained in Example 2 was separated on chiral columnfor the other one of its enantiomers by applying SFC method, and thecorresponding fraction was collected. The solvent was removed by rotaryevaporation to give the pure optical isomer. The preparation conditionsare as follows: chiral column: CHIRALPAK ID; chiral column model: 0.46cm I.D.×15 cm L; sampling quantity: 0.5 μL; mobilephase:CO₂/(EtOH:DCM:NH₄OH=60:40:0.1)=55/45 (V/V); flow rate: 2.0 ml/min;wavelength: UV 254 nm; 103 mg of the isomer B of compound 2 was obtainedwith a retention time of 7.107 min.

The structure of the isomer B was characterized as follows:

¹H NMR (400 MHz, DMSO-d₆) δ 12.35 (brs, 1H), 10.16 (s, 1H), 7.88 (d,J=8.7 Hz, 2H), 7.56 (s, 1H), 7.53 (s, 2H), 7.50 (d, J=1.9 Hz, 1H),7.48-7.44 (m, 2H), 7.39 (d, J=8.7 Hz, 2H), 7.12 (d, J=8.8 Hz, 1H), 6.99(d, J=8.8 Hz, 2H), 4.26 (d, J=14.3 Hz, 1H), 4.16 (d, J=14.4 Hz, 1H),4.13-4.08 (m, 2H), 3.86-3.73 (m, 2H), 3.73-3.68 (m, 2H), 3.46 (t, J=6.5Hz, 4H), 3.30 (s, 2H), 2.38 (s, 2H), 1.66 (q, J=7.4 Hz, 2H), 1.61 (s,3H), 1.55-1.44 (M, 4H), 1.36-1.30 (M, 2H), 1.16 (s, 6H), 0.88 (t, J=7.4Hz, 3H), 0.82 (t, J=7.3 Hz, 3H). ESI-MS (M/z): 755.3 [M+H]⁺.

specific optical rotation: [α]_(280nm) ^(20° C.)=−49.0 (c=0.046, MeOH).

Example 5(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((5-methyl-1-propyl-1H-imidazol-4-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid (Compound 5)

Step I: Synthesis of 5-methyl-1-propyl-1H-imidazol-4-yl-formaldehyde

At room temperature, compound 2-1 (500 mg, 4.5 mmol), potassiumcarbonate (941 mg, 6.8 mmol) and N,N-dimethylformamide (10 mL) wereadded into a three-necked flask, and iodopropane (1.5 g, 9.1 mmol) wasadded thereto. Under the protection of nitrogen gas, the mixture waswarmed to 50° C. and reacted at the temperature for 16 h, and thencooled to room temperature. The system was poured into ice water (100mL) and extracted with ethyl acetate (40 mL×3). The organic phase waswashed with saturated saline (30 mL×3), dried over anhydrous sodiumsulfate, and concentrated. Purification was performed on columnchromatography to give 230 mg of the title compound. ESI-MS (M/z) 153.1[M+H]⁺.

Step II: Synthesis of (5-methyl-1-propyl-1H-imidazol-4-yl) methanol

Compound 5-2 (210 mg, 1.4 mmol) was dissolved in anhydrous methanol (5mL) and cooled to 0° C., and sodium borohydride (66 mg, 1.7 mmol) wasadded thereto in batches. The mixture was reacted at the temperature for30 min. The system was added with water (10 mL) to quench the reactionand concentrated, and ethyl acetate (30 mL×2) was added thereto forextraction. The organic phase was washed with saturated brine (10 mL×2),dried over anhydrous sodium sulfate, and concentrated to give 155 mg ofthe title compound. ESI-MS (M/z): 155.1 [M+H]⁺.

Step III: Synthesis of 4-(chloromethyl)-5-methyl-1-propyl-1H-imidazolehydrochloride

Compound 5-3 (155 mg, 1.0 mmol) was dissolved in acetonitrile (3 mL),and added with thionyl chloride (1 mL) at room temperature, and thenwarmed to 70° C. and reacted for 2 h. The reaction mixture was cooled toroom temperature, concentrated, added with ethyl acetate (5 mL), andstirred at room temperature for 30 minutes, and then suction filtrationwas performed to give 175 mg of the title compound. ESI-MS (M/z) 173.1[M+H]⁺.

Step IV: Synthesis of4-(((5-methyl-1-propyl-1H-imidazol-4-yl)methyl)thio) aniline

At room temperature, 4-aminothiophenol (240 mg, 1.9 mmol), triethylamine(576 mg, 5.7 mmol) and tetrahydrofuran (10 mL) were added to athree-necked flask and cooled to 0° C., and a tetrahydrofuran aqueoussolution (5 mL: 0.5 mL) of compound 5-4 (400 mg, 1.9 mmol) was addeddropwise thereto. After the addition, the mixture was warmed to roomtemperature and reacted for 2 hours. The system was poured into water(50 mL) and extracted with ethyl acetate (40 mL×3). The organic phasewas washed with saturated brine (30 mL×3), dried over anhydrous sodiumsulfate, and concentrated, purification was performed on preparativehigh performance liquid chromatography to give 160 mg of the titlecompound. ESI-MS (M/z) 262.1 [M+H]⁺.

Step V: Synthesis of4-(((5-methyl-1-propyl-1H-imidazol-4-yl)methyl)sulfinyl) aniline

Compound 5-5 (160 mg, 0.61 mmol) was dissolved in dichloromethane (5 mL)and cooled to 0° C., m-chloroperoxybenzoic acid (105 mg, 0.61 mmol) wasadded thereto in batches. After the addition, the mixture was reactedfor 30 min at the temperature. The system was added with saturatedanhydrous sodium sulfite solution (10 mL) to quench the reaction, andextracted by adding dichloromethane (20 mL×3). The organic phase waswashed with saturated brine (20 mL×2), dried over anhydrous sodiumsulfate, and concentrated. Purification was performed on preparativehigh performance liquid chromatography to give 180 mg of the titlecompound. ESI-MS (M/z) 278.1 [M+H]⁺.

Step VI: Synthesis of methyl(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((5-methyl-1-propyl-1H-imidazol-4-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropionate

Compound 5-6 (330 mg, 0.65 mmol) was dissolved in dichloromethane (10mL), and (S)-4-{[(4-methyl-1-propyl-1H-imidazol-5-yl)methyl]sulfinyl}aniline (180 mg, 0.65 mmol) was added to the reaction system.1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (376 mg, 1.95mmol), and 4-dimethylaminopyridine (237 mg, 1.95 mmol) were added andreacted at room temperature overnight. The reaction mixture was addedwith water (20 mL) and extracted with dichloromethane (50 mL×3). Theorganic phases were combined, dried over anhydrous sodium sulfate,concentrated, and purified by column chromatography to give 230 mg ofthe title compound. ES I-MS (M/z) 770.3 [M+H]⁺.

Step VII: Synthesis of(E)-3-(8-(4-(2-butoxyethoxy)phenyl)-5-((4-(((5-methyl-1-propyl-1H-imidazol-4-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid

Compound 5-7 (230 mg, 0.30 mmoL) was dissolved intetrahydrofuran/methanol (10 mL/10 mL), and the lithium hydroxidesolution (5 mL, 1M) was added thereto. The system was warmed to 70° C.and reacted for 2 h. After the reaction was cooled to room temperature,it was adjusted with 1M hydrochloric acid to pH=7, and water (50 mL) wasadded thereto, and extracted with ethyl acetate (30 mL×3). The organicphase was washed with saturated brine (30 mL×2), dried over anhydroussodium sulfate, concentrated, and purified by preparative highperformance liquid chromatography to give 60 mg of the title compound 5.

The structure of compound 5 was characterized as follows:

¹H NMR (400 MHz, Chloroform-d) δ 9.31 (s, 1H), 7.91 (s, 2H), 7.54 (dd,J=14.8, 3.9 Hz, 4H), 7.41 (dd, J=13.6, 6.1 Hz, 4H), 7.02 (s, 2H), 6.93(s, 1H), 4.26 (d, J=15.2 Hz, 1H), 4.20 (s, 2H), 4.10 (d, J=10.4 Hz, 1H),3.89 (s, 2H), 3.85 (s, 2H), 3.59 (s, 2H), 3.42 (s, 2H), 3.14 (s, 2H),2.68 (s, 2H), 2.21 (s, 3H), 1.82 (s, 2H), 1.10 (s, 6H), 1.02-0.97 (M,6H). ESI-MS (M/z): 755.4 [M+H]⁺.

Example 6 Synthesis of(E)-3-(8-(4-(2-methoxyethoxy)phenyl)-5-((4-(((1-(2-methoxyethyl)-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid (Compound 6)

Step I: Synthesis of methyl(E)-8-(4-(2-methoxyethoxy)phenyl)-1,2,3,4-tetrahydrobenzo[b]azacyclooctatetraene-5-formate

Compound 6-1 (816 mg, 2.55 mmoL), 4-(2-methoxyethoxy) phenylboronic acid(500 mg, 2.55 mmoL), palladium acetate (20 mg, 0.085 mmoL), potassiumphosphate (540 mg, 2.53 mmoL), tri-tert-butylphosphoniumtetrafluoroborate (98 mg, 0.35 mmoL), and toluene/water 10:1 (5 mL) wereadded into a microwave tube and reacted under the protection of nitrogengas at the microwave condition of 100° C. for 3 h. After being cooled toroom temperature, the system was poured into water (50 mL) and extractedwith ethyl acetate (30 m×3), and the organic phase was washed withsaturated brine (30 mL×3), dried over anhydrous sodium sulfate,concentrated, and purified by column chromatography to give 530 mg ofthe title compound. ESI-MS (M/z) 410.2 [M+H]⁺.

Step II: Synthesis of(E)-8-(4-(2-methoxyethoxy)phenyl)-1,2,3,4-tetrahydrobenzo[b]azacyclooctatetraene-5-formicacid

Compound 6-2 (530 mg, 1.45 mmoL) was dissolved intetrahydrofuran/methanol (10 mL/10 mL), and sodium hydroxide solution (5mL, 1M) was added. The system was heated to 70° C. and reacted for 2 h.After the reaction was cooled to room temperature, it was adjusted with1M hydrochloric acid to pH=7. The system was added with water (50 mL),and extracted with ethyl acetate (30 mL×3). The organic phase was washedwith saturated brine (30 mL×2), dried over anhydrous sodium sulfate,concentrated, and purified by column chromatography to give 388 mg ofthe title compound. ES I-MS (M/z) 396.2 [M+H]⁺.

Step III: Synthesis of(E)-8-(4-(2-methoxyethoxy)phenyl)-1-(3-methoxy-2,2-dimethyl-3-oxopropyl)-1,2,3,4-tetrahydrobenzo[b]azacyclooctatetraene-5-formicacid

Oxalyl chloride (3.818 g, 30 mmol) was dissolved in dichloromethane andcooled to −78° C., and then under the protection of nitrogen gas,dimethyl sulfoxide (3.12 g, 40 mmol) was added dropwise thereto. Themixture was reacted at the temperature for 30 min. The above reactionliquid was added with dichloromethane solution of methyl2,2-dimethyl-3-hydroxypropionate (1.32 g, 10 mmol), and reacted at thetemperature for 2 h. Triethylamine (10 g, 100 mmol) was added theretoand the mixture was reacted for 1 hour at the temperature. The mixturewas warmed to −20° C., and trifluoroacetic acid was added dropwise toallow the reaction system to be acidic (pH=1.0). Compound 6-3 (388 mg,1.1 mmol) and sodium triacetoxyborohydride (2.2 g, 10 mmol) were addedto the reaction system, and the resulting mixture was reacted at thetemperature for 1 hour. The reaction solution was added with distilledwater (100 mL), and extracted with dichloromethane (30 mL×3). Theorganic phases were combined and washed with saturated sodiumbicarbonate (100 mL×1), saturated brine (100 mL×1), and water (100 mL×1)successively, dried over anhydrous sodium sulfate, and purified bycolumn chromatography to give 300 mg of the title compound. ES I-MS(M/z) 510.3 [M+H]⁺.

Step IV: Synthesis of methyl(E)-3-(8-(4-(2-methoxyethoxy)phenyl)-5-((4-(((1-(2-methoxyethyl)-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropionate

Compound 6-4 (300 mg, 0.65 mmol) was dissolved in dichloromethane (10mL), and (S)-4-{[(1-(2-methoxyethyl)-1H-imidazol-5-yl)methyl]sulfinyl}aniline (180 mg, 0.65 mmol) was added to the reaction system, followedby an addition of 1-ethyl-(3-dimethylaminopropyl)carbodiimidehydrochloride (376 mg, 1.95 mmol) and 4-dimethylaminopyridine (237 mg,1.95 mmol), then reacted at room temperature overnight. The reactionmixture was added with water (20 mL) and extracted with dichloromethane(50 mL×3). The organic phases were combined, dried over anhydrous sodiumsulfate, concentrated, and purified by column chromatography to give 155mg of the title compound. ESI-MS (M/z) 771.3 [M+H]⁺.

Step V: Synthesis of(E)-3-(8-(4-(2-methoxyethoxy)phenyl)-5-((4-(((1-(2-methoxyethyl)-1H-imidazol-5-yl)methyl)sulfinyl)phenyl)carbamoyl)-3,4-dihydrobenzo[b]azacyclooctatetraen-1(2H)-yl)-2,2-dimethylpropanoicacid

Compound 6-5 (155 mg, 0.20 mmoL) was dissolved intetrahydrofuran/methanol (10 mL/10 mL), and sodium hydroxide solution (5mL, 1M) was added thereto. The system was heated to 70° C. and reactedfor 2 h. After the reaction was cooled to room temperature, the systemwas adjusted with 1M hydrochloric acid to pH=7. The system was addedwith water (50 mL) and extracted with ethyl acetate (30 mL×3). Theorganic phase was washed with saturated brine (30 mL×2), dried overanhydrous sodium sulfate, concentrated, and purified by preparative highperformance liquid chromatography to give 8 mg of the title compound 6.

The structure of compound 6 was characterized as follows:

¹H NMR (400 MHz, Chloroform-d) δ 9.37 (s, 1H), 8.29 (s, 1H), 7.96 (d,J=8.3 Hz, 2H), 7.57 (dd, J=8.4, 2.2 Hz, 1H), 7.52 (d, J=8.2 Hz, 2H),7.41 (s, 1H), 7.37 (d, J=8.5 Hz, 1H), 7.28 (d, J=8.3 Hz, 2H), 7.03 (d,J=8.3 Hz, 2H), 6.88 (s, 1H), 6.58 (s, 1H), 4.37 (d, J=14.6 Hz, 1H),4.32-4.25 (m, 1H), 4.20 (t, J=4.9 Hz, 2H), 4.11 (d, J=15.7 Hz, 1H), 3.95(d, J=14.7 Hz, 1H), 3.85 (t, J=5.0 Hz, 2H), 3.64 (d, J=8.4 Hz, 2H), 3.59(d, J=6.7 Hz, 2H), 3.44 (d, J=15.9 Hz, 2H), 3.34 (s, 3H), 3.15 (s, 2H),2.68 (d, J=7.6 Hz, 2H), 1.65 (t, J=7.5 Hz, 4H), 1.46-1.42 (m, 2H), 1.13(s, 3H), 1.09 (s, 3H), 0.98 (d, J=7.4 Hz, 3H), ESI-MS (m/z): 757.4[M+H]⁺.

Pharmacological Activity Assay

The following examples further illustrate the invention, but are notmeant to limit the scope of the invention.

Experimental Example 1: Inhibition Test on Activity of CCR2/CCR5 inCells

Test Principles

HEK293 cells that stably express CCR2 and CCR5 receptor proteins werefirst labeled with calcium ion sensitive fluorescent probe and thenstimulated with corresponding ligands (MCP1 and RANTES). After thestimulation, the receptors were activated to trigger the release of thecalcium ions from endoplasmic reticulum to cytoplasm. And once thecalcium ions were captured by the fluorescent probe, the fluorescentsignal could be triggered. The inhibition of compounds on CCR2 and CCR5can be evaluated by detecting the intensity of the fluorescent signal.

Materials Used for Experiments:

HEK293 cells: WuXi Apptec Co., Ltd;

384-well plates (384 well Poly-D-Lysine protein coating plate), Greiner#781946;

Monocyte chemotactic protein 1 (MCP 1): CCR2 Agonist, PeproTech-300-04;

Regulating and activating normal T-cell expression secretion factor(RANTES): CCR5 Agonist, PeproTech-300-06.

To each well of the 384-well plates, 2×10⁴ HEK293 cells were added andcultured overnight. After being labelled by fluorescent probe, MCP1 andRANTES in different concentrations were added, and the fluorescentsignal was read. The fluorescence signal was plotted in relation to theligand concentration, and according to the four-parameter model, IC₅₀values were calculated as the ligand concentrations of the CCR2 and CCR5for the subsequent detection of the compounds.

By referring to the above results, cells were incubated with the testcompound in different concentrations (5000 nM, 1250 nM, 313 nM, 78 nM,20 nM, 4.8828 nM, 1.2207 nM, 0.3052 nM, 0.0763 nM, and 0.0191 nM), andafter MCP1 and RANTES were added thereto, the fluorescent signal wasread. The inhibition activity of each concentration group wascalculated, wherein the solvent group (not containing the compound) wasas the negative control, and the Buffer group (not containing anyligand) was as the blank control:

Relative inhibition activity percentage=1−(fluorescent signal of eachconcentration group-fluorescent signal of the blank group)/(fluorescentsignal of the solvent group-fluorescent signal of the blank group)*100%.

The inhibition activity percentage of each concentration group wasplotted in relation to the concentration of the compound, and the curvewas fitted according to the four-parameter model, IC₅₀ values werecalculated:

y=min+(max−min)/(1+(x/IC₅₀){circumflex over ( )}(−Hillslope))

wherein y is the relative inhibition activity percentage; max, min arethe maximum and minimum values of the fitted curve, respectively; x isthe concentration of the compound, and Hillslope is the slope of thecurve.

Test Results

According to the above method, the inhibition activity of the compoundagainst CCR2/CCR5 was determined, and the results are shown in Table 1:

TABLE 1 Inhibition results of the compound against the CCR2/CCR5 incells IC₅₀ (nM) Compound No. CCR2 CCR5 Compound 1 0.74 0.33 Compound 26.38 3.18 Compound 3 0.81 1.48 Compound 4 6.12 2.82

As can be seen from Table 1, the compounds of the present application(e.g. compounds 1, 2, 3 and 4) have a extremely potent inhibitory effectagainst CCR2/CCR5.

Experimental Example 2: Inhibition Tests Against CYP 1A2, CYP 2D6, CYP3A4M, CYP 3A4T and hERG 1. Inhibition Tests Against CYP 1A2, CYP 2D6,CYP 3A4M, and CYP 3A4T

CYP 1A2:

Probe substrate: Phenacetin, National Institute of Control ofPharmaceutical & Biological Products, 100095.

Positive control: alpha-naphthoflavane, ACras, A0331533.

CYP 2D6:

Probe substrate: Dextromethorpha, Beijing Jinbao, 100201.

Positive control: Quinidine, Damas-bata, P1176507.

CYP 3A4M:

Probe substrate: Midazolam, National Institute of Control ofPharmaceutical & Biological Products, 171265.

Positive control: Ketoconazole, purchased from Dingdang Chemical, DTYC0516-17.

CYP 3A4T:

Probe substrate: Testosterone, European Pharmacopoeia (EP) ReferenceStandard, Sigma-Aldrich, T0100000.

Positive control: Ketoconazole, Dingdang Chemical, DTYC 0516-17.

A probe substrate (50 μl), PBS (49 μl) and a test compound in differentconcentrations (0.05 μM, 0.15 μM, 0.5 μM, 1.5 μM, 5 μM, 15 μM, 50 μM) ora positive control (1 μl) were mixed with human liver microsomes (HLM,50 μl) to give 150 μl of a mixed liquid, which was subjected topre-incubation (37° C.) for 5 min, then added with NADPH (50 μl) andincubated for 30 min. The reaction was quenched by adding 200 μl ofglacial acetonitrile. An appropriate volume of an internal standardsubstance was added, and after vortex treatment and centrifugation, thesupernatant was obtained.

The detection was performed by using LC-MS/MS. The mass spectrometer wasAPI 5500, the liquid chromatograph was Waters ACQUITY UPLC I-CLASSsystem. The chromatographic column was Hypersil GOLD C18, 1.9 μmparticle size, 50×2.1 mm; mobile phase A: water+0.1% formic acid, mobilephase B: acetonitrile; flow rate: 0.4 ml/min, column temperature: 40° C.The ion source as used was ESI source positive ion mode, and thescanning mode was Multiple Reaction Monitoring (MRM).

By using solvent group (DMSO) as negative control, the concentrations ofthe main metabolite converted from the probe substrate in the case ofdifferent concentrations of the test compound were measured, todetermine the half inhibition concentration of the compound (IC₅₀). Thehalf inhibition concentration (IC₅₀) results of the compound were shownin Tables 2-3.

2. hERG Inhibition Test:

Different concentrations (1 μM and 10 μM) of the test compound and 30 μMpositive control Quinidine were formulated. HEK293 cells over-expressinghERG potassium ion channels were cultured in 5% CO₂ incubator at 37° C.When the density of the cells reaches 80% of the Petri dish, the cellswas washed with phosphate buffer solution (PBS), followed by digestionwith Trypsin/EDTA for 2-3 minutes. The digestion was terminated byadding cell culture medium, then the solution was transferred andcentrifuged to remove the supernatant, and added with cell culturemedium (DMEM, 15% fetal bovine serum and 1%100×Penicillin-Streptomycin). After uniformly mixing, the cells weredripped onto a circular slide and cultured in a Petri dish to obtain thetest cells.

Channel current was recorded by applying electrophysiological manualpatch clamp in a whole-cell recording mode: extracellular fluid wasperfused until the current was stable, after the peak value of the tailcurrent (control current value) was recorded, the extracellular fluidcontaining different concentrations of the test compound was perfused,continuously recording was conducted until the inhibition effect of themedicine on the hERG current reach stable, and the peak value of thetail current (current value after drug administration) was recorded.

The results of this experiment were shown in Table 4.

TABLE 2 Inhibition results of the compounds against CYP CYP 1A2 CYP 2D6Compound (μM) (μM) Compound 1 >10 >10 Compound 2 >10 >10 Compound3 >10 >10 Compound 4 >10 >10

TABLE 3 Inhibition results of the compounds against CYPP CYP 3A4M CYP3A4T Compound (μM) (μM) Cenicriviroc 3.43 12.7 Compound 2 27.8 62.2Compound 3 13.3 32.3 Compound 4 15.3 48.3

TABLE 4 Inhibition results of the compounds against hERG Compound hERG(μM) Compound 2 >10 Compound 3 >10 Compound 4 >10

As can be seen from Tables 2-4, compounds of the present applicationhave weak induction effect on CYP enzyme, no inhibition effect on hERGchannel, and no obvious cardiotoxicity. The compounds of the presentapplication (such as compound 1, compound 2, compound 3 and compound 4)have better properties in the aspects of CYP and hERG and have bettersafety.

Experimental Example 3: Rat Pharmacokinetic (PK) Studies

The test compound was administered intragastrically (p.o.) to male SDrats at a dose of 5 mg/kg. The solvent system was 0.5% MC during thep.o. administration. After p.o. administration, blood was collected forPK studies at 5 min, 15 min, 30 min, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h.After p.o. administration, liver tissues were collected at 15 min, 1 h,4 h, 8 h and 24 h, and the liver tissue sample, after being subjected toprotein precipitation treatment, was analyzed by a LC-MS/MS. The massspectrometer was API 5500, and the liquid chromatograph was WatersACQUITY I CLASS system: chromatographic column: Agela ASB C18 column(2.1 mm×50 mm, 1.9 μm); mobile phase A: water+0.1% formic acid, mobilephase B: acetonitrile; flow rate: 0.4 mL/min, column temperature: 40° C.The ion source as used was ESI source positive ion mode, and thescanning mode was Multiple Reaction Monitoring (MRM). The test resultswere shown in Table 5.

TABLE 5 Rat pharmacokinetic data of the compound Admini- Compoundstration Dose No. Route Object (mg/kg) AUC_(last) C_(max) T_(1/2) (h) F% Compound p.o. Blood 5.00 165 ± 36 49.7 ± 10.6 1.93 ± 0.37 20.9 ± 4.5 3plasma (h * ng/ml) (ng/ml) Liver 5.00 386938 ± 30242 26600 ± 200 7.20 ±0.50 / (h * ng/g) (ng/g) Cenicriviroc p.o. Blood 5.00 6326 ± 3440 746 ±428 4.33 ± 0.87 10.8 ± 3.5 plasma (h * ng/ml) (ng/ml) Liver 5.00 6033 ±2193 688 ± 229 3.93 ± 1.88 / (h * ng/g) (ng/g) “/” indicates absence

As can be seen from Table 5, the exposure of the compound of the presentapplication (for example compound 3) in the liver of the rats was about2345 folds higher than that in the plasma; the drug concentration in theliver was about 530 folds higher than that in the plasma; the T_(1/2) inthe liver was 7.20±0.50 h. The compound of the present application hadvery low exposure quantity (AUC_(last)) and drug concentration (C_(max))in the plasma, whereas had very high exposure quantity and drugconcentration in the liver, and thus it has very remarkable livertargeting property and good pharmacokinetic properties.

The other compounds in the present application also had similarAUC_(last), C_(max) and T_(1/2) to the above compound, and they hadsimilar pharmacokinetic properties in rats to those as above described.

In summary, the compound of the present application shows goodactivities in the aspect of in vitro drug efficacy, and have excellentpharmacokinetic properties and remarkable liver targeting property, andthus they can be applied in the manufacture of drugs for treatment ofCCR2- and CCR5-mediated diseases, especially nonalcoholic fatty liverdiseases (NAFLD) or the like.

In addition to those embodiments as described herein, according to thepreceding depictions, many modifications to the invention are obviousfor a person skilled in the art. Such modifications are also intended tofall within the scope of the claimed as attached. Each referencedocuments (including all patents, patent applications, journals, books,and any other publications) cited in the present application areincorporated as the reference in its entirety.

1. A compound of general formula I or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned:

wherein R¹ is selected from hydrogen, deuterium, and C₁₋₆ alkyl, saidC₁₋₆ alkyl is optionally substituted with R²; L₁ is selected from C₁₋₆alkylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and3- to 10-membered heterocyclylene; L₂ is absent or selected from —NH—,C₂₋₆ alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, C₁₋₆ alkyl, C₁₋₆haloalkyl, alkoxy, C₁₋₆ haloalkoxy, —C(O)C₁₋₄ alkyl, —COOR³, —N(R³)₂,C₃₋₁₀ cycloalkyl, 3- to 10-membered heterocyclic group, C₆₋₁₀ aryl,—CON(R³)₂ and —NR³CO₂R³; said 3- to 10-membered heterocyclylene containsone or more ring members selected from N, NR³, O, P, and S(O)_(z);wherein y₁ and y₂ are each independently selected from 0, 1, 2, 3 or 4;z is selected from 0, 1 or 2; L³ is absent or selected from C₁₋₆alkylene, C₂₋₆ alkenylene, C₂₋₆ alkynylene, C₃₋₁₀ cycloalkylene, C₆₋₁₀arylene, C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene; R²is selected from hydrogen, deuterium, hydroxyl, amino, carboxyl, C₁₋₆alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆ alkyleneoxy)_(m2)-C₁₋₆ alkyl,(C₁₋₆ alkyl)₂ amino, and 5- to 6-membered heterocyclyl containing one ormore N, O or S; wherein m₁, m₂ are each independently selected from 0,1, 2, 3 or 4; R³ is selected from hydrogen, deuterium, C₁₋₄ alkyl andC₃₋₆ cycloalkyl; A is selected from 4- to 7-membered heterocyclyl or 5-to 10-membered heteroaryl, both of which are optionally substituted witha substituent selected from deuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkylthat are optionally substituted with R⁶, halogen, —CN, halogenated C₁₋₆alkyl, C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂ heteroaryl; R⁶ isselected from deuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆cycloalkoxy; n is selected from 0, 1 or
 2. 2. The compound according toclaim 1 or a pharmaceutically acceptable salt, ester, solvate (e.g.,hydrate), stereoisomer, tautomer, prodrug thereof, or crystalline form,metabolite thereof, or a mixture of the aforementioned, wherein, R¹ isselected from hydrogen and C₁₋₄ alkyl, said C₁₋₄ alkyl is optionallysubstituted with R²; R² is selected from hydroxyl, carboxyl, C₁₋₆alkoxy, —(C₁₋₆ alkyleneoxy)_(m1)-(C₁₋₆ alkyleneoxy)_(m2)-C₁₋₆ alkyl and5- to 6-membered heterocyclyl containing one or more N, O or S; whereinm₁ and m₂ are independently selected from 0, 1, 2, 3 or 4; preferably,R¹ is selected from C₁₋₄ alkyl, said C₁₋₄ alkyl is optionallysubstituted with R²; R² is selected from hydroxyl, C₁₋₆ alkoxy, —(C₁₋₆alkyleneoxy)_(m1)-(C₁₋₆ alkyleneoxy)_(m2)-C₁₋₆ alkyl and 5- to6-membered heterocyclyl containing one or more N, O or S; wherein m₁ andm₂ are each independently selected from 0, 1 or 2; preferably, R¹ isselected from C₁₋₄ alkyl, said C₁₋₄ alkyl is optionally substituted withR²; R² is selected from C₁₋₄ alkoxy, —(C₁₋₄ alkyleneoxy)_(m1)-(C₁₋₄alkyleneoxy)_(m2)-C₁₋₄ alkyl and 5- to 6-membered heterocyclylcontaining one or more N, O or S; wherein m₁ and m₂ are eachindependently selected from 0, 1 or 2; preferably, R¹ is selected fromC₁₋₄ alkyl, said C₁₋₄ alkyl is optionally substituted with R²; R² isselected from C₁₋₄ alkoxy; preferably, R¹ is


3. The compound according to claim 1 or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein, L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, and C₅₋₁₂ heteroarylene; preferably, L₁ isselected from C₁₋₆ alkylene, C₃₋₇ cycloalkylene, C₆₋₈ arylene, and C₅₋₈heteroarylene; preferably, L₁ is selected from C₁₋₃ alkylene and C₃₋₅cycloalkylene; preferably, L₁ is selected from —CH₂— or —CH₂—CH₂—. 4.The compound according to claim 1 or a pharmaceutically acceptable salt,ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof,or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein, L₂ is absent or selected from —NH—, C₂₋₆alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂ heteroarylene, and 3- to 10-memberedheterocyclylene; said —NH—, C₂₋₆ alkenylene, —(C₁₋₆alkyleneoxy)_(y1)-(C₁₋₆ alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀arylene, C₅₋₁₂ heteroarylene, and 3- to 10-membered heterocyclylene areoptionally substituted with one or more substituents independentlyselected from hydrogen, deuterium, halogen, hydroxyl, cyano, C₁₋₆ alkyl,C₁₋₆ haloalkyl, C₁₋₆ alkoxy, C₁₋₆ haloalkoxy, —C(O)C₁₋₄ alkyl, —COORS,—N(R³)₂, C₃₋₁₀ cycloalkyl, 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl,—CON(R³)₂ and —NR³CO₂R³; said 3- to 10-membered heterocyclylene containsone or more ring members selected from N, NR³, O, P, and S(O)_(z);wherein y₁ and y₂ are each independently selected from 0, 1, 2, 3 or 4;z is selected from 0, 1 or 2; preferably, L₂ is absent or selected from—NH—, C₂₋₆ alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, C₁₋₃ alkyl, C₁₋₃haloalkyl, C₁₋₃ haloalkoxy, —C(O)C₁₋₄ alkyl, —COORS and —NR³CO₂R³; said3- to 10-membered heterocyclylene contains one or more ring membersselected from N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂ are eachindependently selected from 0, 1, 2, 3 or 4; z is selected from 0, 1 or2; R³ is selected from hydrogen, deuterium, C₁₋₄ alkyl and C₃₋₆cycloalkyl; preferably, L₂ is absent or selected from —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene; said —NH—, C₂₋₆alkylene, C₂₋₆ alkenylene, —(C₁₋₆ alkyleneoxy)_(y1)-(C₁₋₆alkyleneoxy)_(y2)-, C₃₋₁₀ cycloalkylene, C₆₋₁₀ arylene, C₅₋₁₂heteroarylene, and 3- to 10-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, methyl, halomethyl,halomethoxy, —C(O)CH₃, —COOR³ and —NR³CO₂R³; said 3- to 10-memberedheterocyclylene contains one or more ring members selected from N, NR³,O, P, and S(O)_(z); wherein y₁ and y₂ are each independently selectedfrom 0, 1, 2, 3 or 4; z is selected from 0, 1 or 2; R³ is selected fromhydrogen, deuterium, methyl and cyclopropyl; preferably, L₂ is absent orselected from —NH—, C₂₋₄ alkylene, C₂₋₃ alkenylene, —(C₁₋₃alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈arylene, C₅₋₈ heteroarylene, and 3- to 6-membered heterocyclylene; said—NH—, C₂₋₄ alkylene, C₂₋₃ alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene,C₅₋₈-heteroarylene, and 3- to 6-membered heterocyclylene are optionallysubstituted with one or more substituents independently selected fromhydrogen, deuterium, halogen, hydroxyl, cyano, methyl, halomethyl,halomethoxy, —C(O)CH₃, —COOR³ and —NR³CO₂R³; said 3- to 6-memberedheterocyclylene contains one or more ring members selected from N, NR³,O, P, and S(O)_(z); wherein y₁, y₂ are each independently selected from0, 1, 2, 3 or 4; z is selected from 0, 1 or 2; R³ is selected fromhydrogen, deuterium, methyl and cyclopropyl; preferably, L₂ is absent orselected from —NH—, —N(CH₃)—, —C(CH₃)₂—, —CH═CH—, —(C₁₋₃alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈arylene, C₅₋₈ heteroarylene, and 3- to 6-membered heterocyclylene,wherein y₁ and y₂ are each independently selected from 0, 1, or 2;preferably, L₂ is absent or selected from —NH—, —N(CH₃)— and —C(CH₃)₂—.5. The compound according to claim 1 or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein, L₃ is absent or selected from C₁₋₃ alkylene,C₂₋₃ alkenylene, C₂₋₃ alkynylene, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 7-membered heterocyclylene; preferably, L₃ isabsent or selected from C₁₋₆ alkylene and C₂₋₆ alkenylene; preferably,L₃ is absent or selected from —C(CH₃)₂—, —CH₂—, and —CH═CH—; preferably,L₃ is absent or —CH₂—.
 6. The compound according to claim 1 or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned, wherein, A is selected from4- to 7-membered heterocyclyl or 5- to 10-membered heteroaryl;preferably, A is selected from 4- to 7-membered nitrogen-containingheterocyclyl or 5- to 10-membered nitrogen-containing heteroaryl;preferably, A is selected from 4- to 7-membered nitrogen-containingheterocyclyl or 5- to 10-membered nitrogen-containing heteroaryl, bothof which are optionally substituted with a substituent selected fromhydrogen, deuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl that are optionallysubstituted with R⁶, halogen, —CN, halogenated C₁₋₆ alkyl, C₃₋₆heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂ heteroaryl; R⁶ is selected fromdeuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆ cycloalkoxy; preferably,A is selected from 5- or 6-membered nitrogen-containing heteroaryl;preferably, A is selected from 5- or 6-membered nitrogen-containingheteroaryl optionally substituted with a substituent selected fromdeuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl that are optionallysubstituted with R⁶, halogen, —CN, halogenated C₁₋₆ alkyl, C₃₋₆heterocycloalkyl, C₆₋₁₀ aryl and C₅₋₁₂ heteroaryl; R⁶ is selected fromdeuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆ cycloalkoxy; preferably,A is a 5-membered nitrogen-containing heteroaryl; preferably, A is a5-membered nitrogen-containing heteroaryl optionally substituted with asubstituent selected from deuterium, C₁₋₆ alkyl and C₃₋₆ cycloalkyl thatare optionally substituted with R⁶, halogen, —CN, halogenated C₁₋₆alkyl, C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryl, and C₅₋₁₂ heteroaryl; R⁶ isselected from deuterium, hydroxyl, —CN, C₁₋₆ alkoxy and C₃₋₆cycloalkoxy; preferably, A is a 5-membered nitrogen-containingheteroaryl substituted with a substituent, and the substituent is C₁₋₆alkyl (e.g., propyl).
 7. The compound according to claim 6 or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned, wherein A is

wherein R⁴ is selected from hydrogen, deuterium, or C₁₋₆ alkyl and C₃₋₆cycloalkyl that are optionally substituted with R⁶, R⁶ is selected fromdeuterium, hydroxyl, —CN, C₁₋₄ alkoxy and C₃₋₆ cycloalkoxy; preferably,R⁴ is C₁₋₆ alkyl optionally substituted with R⁶, R⁶ is selected fromdeuterium, hydroxyl, —CN, and C₁₋₄ alkoxy; preferably, R⁴ is C₁₋₆ alkyloptionally substituted with R⁶, R⁶ is selected from hydroxyl and C₁₋₄alkoxy; preferably, R⁴ is C₁₋₄ alkyl optionally substituted with R⁶, R⁶is selected from hydroxyl and C₁₋₄ alkoxy; preferably, R⁴ is C₁₋₄ alkyloptionally substituted with R⁶, R⁶ is selected from hydroxyl, methoxy,and ethoxy; preferably, R⁴ is C₁₋₄ alkyl optionally substituted with R⁶,R⁶ is selected from methoxy and ethoxy; preferably, R⁴ is selected fromn-propyl and

each R⁵ is selected from hydrogen, deuterium, halogen, —CN, C₁₋₆ alkyl,halogenated C₁₋₆ alkyl, C₁₋₆ alkylene-CN, C₁₋₆ alkylene-OH, C₁₋₆alkylene-C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, C₃₋₆ heterocycloalkyl, C₆₋₁₀ aryland C₅₋₁₂ heteroaryl; preferably, each R⁵ is independently selected fromhydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl, C₁₋₆alkylene-OH, C₁₋₆ alkylene-C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₃₋₆heterocycloalkyl; preferably, each R⁵ is independently selected fromhydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl, and C₃₋₆cycloalkyl; preferably, each R⁵ is independently selected from hydrogen,methyl, trifluoromethyl and cyclopropyl; preferably, R⁵ is hydrogen; Xis selected from N and C—R⁵, each R⁵ is identical or different. 8.(canceled)
 9. The compound according to claim 7 or a pharmaceuticallyacceptable salt, ester, solvate (e.g., hydrate), stereoisomer, tautomer,prodrug thereof, or crystalline form, metabolite thereof, or a mixtureof the aforementioned, wherein X is selected from N and C—R⁵, R⁵ isselected from hydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl,C₁₋₆ alkylene-OH, C₁₋₆ alkylene-C₁₋₆ alkoxy, C₃₋₆ cycloalkyl, and C₃₋₆heterocycloalkyl; preferably, X is selected from N and C—R⁵, R⁵ isselected from hydrogen, deuterium, C₁₋₆ alkyl, halogenated C₁₋₆ alkyl,and C₃₋₆ cycloalkyl; X is selected from N and C—R⁵, R⁵ is selected fromhydrogen, methyl, trifluoromethyl, and cyclopropyl.
 10. The compoundaccording to claim 1 or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned, wherein A is selected from


11. The compound according to claim 1 or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein L₁ is selected from C₁₋₆ alkylene, C₃₋₁₀cycloalkylene, C₆₋₁₀ arylene and C₅₋₁₂ heteroarylene; preferably, L₁ isselected from C₁₋₃ alkylene and C₃₋₅ cycloalkylene; more preferably, L₁is selected from methylene and ethylene; L₂ is absent or selected from—NH—, C₂₋₄ alkylene, C₂₋₃ alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃alkyleneoxy)_(y2)-, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroaryleneand 3- to 6-membered heterocyclylene; said —NH—, C₂₋₄ alkylene, C₂₋₃alkenylene, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-, C₃₋₆cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene, and 3- to 6-memberedheterocyclylene are optionally substituted with one or more substituentsthat are independently selected from hydrogen, deuterium, halogen,hydroxyl, cyano, methyl, halomethyl, halomethoxy, —C(O)CH₃, —COOR³ and—NR³CO₂R³; said 3- to 6-membered heterocyclylene contains one or morering members selected from N, NR³, O, P, and S(O)_(z); wherein y₁ and y₂are each independently selected from 0, 1, 2, 3 or 4; z is selected from0, 1 or 2; R³ is selected from hydrogen, deuterium, methyl andcyclopropyl; preferably, L₂ is absent or selected from —NH—, —N(CH₃)—,—C(CH₃)₂—, —CH═CH—, —(C₁₋₃ alkyleneoxy)_(y1)-(C₁₋₃ alkyleneoxy)_(y2)-,C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈ heteroarylene and 3-to 6-memberedheterocyclylene, wherein y₁ and y₂ are each independently selected from0, 1 or 2; more preferably, L₂ is absent or selected from —NH—, —N(CH₃)—and —C(CH₃)₂—; L₃ is absent or selected from C₁₋₃ alkylene, C₂₋₃alkenylene, C₂₋₃ alkynylene, C₃₋₆ cycloalkylene, C₆₋₈ arylene, C₅₋₈heteroarylene, and 3- to 7-membered heterocyclylene; preferably, L₃ isabsent or selected from C₁₋₆ alkylene and C₂₋₆ alkenylene; morepreferably, L₃ is absent or selected from —C(CH₃)₂—, —CH₂—, and —CH═CH—;particularly preferably, L₃ is absent or —CH₂—.
 12. The compoundaccording to claim 1 or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned, wherein the compound has a structure shown by formulaI_(a) or formula I_(b):

wherein R¹, R⁴, R⁵, L₁, L₂, L₃, X and n are as defined in claim
 1. 13.The compound according to claim 1 or a pharmaceutically acceptable salt,ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof,or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein the compound has a structure shown by formulaII_(a) or formula II_(a′):

wherein R¹, R⁴, R⁵, X, L₁, L₂, L₃ and n are as defined in claim
 1. 14.The compound according to claim 1 or a pharmaceutically acceptable salt,ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof,or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein the compound has a structure as shown by formulaII_(b) or formula II_(b′):

wherein R¹, R⁴, R⁵, X, L₁, L₂, L₃ or n are as defined in claim
 1. 15.The compound according to claim 13 or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein n is selected from 1 or 2, preferably, n is 1.16. The compound according to claim 1 or a pharmaceutically acceptablesalt, ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrugthereof, or crystalline form, metabolite thereof, or a mixture of theaforementioned, wherein the compound is selected from:


17. A pharmaceutical composition, comprising at least one compoundaccording to claim 1 or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned, and one or more pharmaceutically acceptable carriers;preferably, the pharmaceutical composition further comprises one or moreother drugs that prevent or treat a CCR2- and/or CCR5-mediated diseaseor condition, especially nonalcoholic fatty liver disease (NAFLD) or thelike; preferably, the pharmaceutical composition is in a form selectedfrom tablets, capsules, lozenges, hard candies, powder, sprays, creams,ointments, suppositories, gels, pastes, lotions, ointments, aqueoussuspensions, injectable solutions, elixirs and syrups. 18.-20.(canceled)
 21. A kit product, comprising: a) a first container,comprising, as a first therapeutic agent, at least one compoundaccording to claim 1 or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned; b) an optional second container, comprising, as a secondtherapeutic agent, at least one other therapeutic agent, or as a secondpharmaceutical composition, a pharmaceutical composition comprising saidother therapeutic agent; and c) an optional packaging specification. 22.(canceled)
 23. A method for the prevention or treatment of a CCR2-and/or CCR5-mediated disease or condition, comprising administering toan individual in need a therapeutically effective amount of the compoundaccording to claim 1 or a pharmaceutically acceptable salt, ester,solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof, orcrystalline form, metabolite thereof, or a mixture of theaforementioned; preferably, the CCR2- and/or CCR5-mediated disease orcondition is preferably nonalcoholic fatty liver disease (NAFLD) or thelike. 24.-25. (canceled)
 26. A process for the preparation of thecompound of formula I according to claim 1, comprising the followingsynthetic route:

wherein R¹, n, L₁, L₂, L₃ and A are as defined in claim 1, L is a groupbearing an aldehyde group at the end, and L may be converted to L₁(preferably, L₁ is C₁₋₃ alkylene, and more preferably, L₁ is selectedfrom methylene and ethylene) after reactions, and the carboxylprotection group PG is selected from the following groups that areoptionally substituted with one or more R⁷: C₁₋₆ alkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, C₃₋₁₀ cycloalkyl, C₃₋₁₀ heterocycloalkyl, C₆₋₁₀ aryl, C₁₋₆alkylene-C₆₋₁₀ aryl, C₅₋₁₂ heteroaryl, R⁷ is selected from hydrogen,deuterium, halogen, C₁₋₆ alkyl and C₁₋₆ alkoxy; preferably, the carboxylprotection group PG is preferably C₁₋₆ alkyl; particularly, the processcomprises the following steps: step I: reacting compound a with compoundb to provide compound c: the reaction is carried out in a suitableorganic solvent, and said organic solvent may be selected from a linearor cyclic ether (e.g. tetrahydrofuran or diethyl ether), a halogenatedhydrocarbon (e.g. dichloromethane, chloroform, 1,2-dichloroethane or thelike), a nitrile (e.g. acetonitrile or the like), N-methylpyrrolidone,dimethylformamide, dimethylacetamide, 1,4-dioxane, dimethylsulfoxide andany combination thereof, preferably the organic solvent is dioxane or1,2-dichloroethane; the reaction is carried out in the presence of asuitable reducing agent, and said reducing agent is selected from sodiumborohydride, potassium borohydride, sodium cyanoborohydride and sodiumborohydride acetate, preferably the reducing agent is sodiumcyanoborohydride; the reaction is carried out under suitable acidiccondition, and said acid may be selected from hydrochloric acid, aceticacid and trifluoroacetic acid, preferably the acid is trifluoroaceticacid; the reaction is preferably carried out at a suitable temperature,and said temperature is preferably from −50 to 100° C., more preferablyfrom −20 to 70° C.; the reaction is carried out for a suitable time, forexample 1 to 24 hours, and for example 1 to 6 hours; Step II: reactingcompound c with compound d to provide compound e: the reaction iscarried out in a suitable organic solvent, and said organic solvent maybe selected from a halogenated hydrocarbon (e.g., dichloromethane,chloroform, 1,2-dichloroethane or the like), a nitrile (e.g.,acetonitrile or the like), N-methylpyrrolidone, dimethylformamide,dimethylacetamide, 1,4-dioxane, dimethylsulfoxide, and any combinationthereof, preferably the organic solvent is dichloromethane; the reactionis carried out in the presence of a suitable condensing agent, and thecondensing agent may be selected from thionyl chloride, oxalyl chloride,phosphorus oxychloride, phosphorus trichloride, phosphoruspentachloride, ethyl chloroformate, isopropyl chloroformate, HATU, HBTU,EEDQ, DEPC, DCC, DIC, EDC, BOP, PyAOP or PyBOP, preferably thecondensing agent is thionyl chloride, oxalyl chloride; the reaction iscarried out under suitable basic condition; the base is an organic orinorganic base; preferably, the organic base is selected fromtriethylamine, DIPEA, pyridine, NMM or DMAP, and preferably, theinorganic base is selected from NaH, NaOH, Na₂CO₃ or K₂CO₃; preferably,the base is selected from triethylamine, DIPEA and DMAP; the reaction iscarried out at a suitable temperature, and the reaction temperature ispreferably from 0 to 100° C., more preferably from 15 to 50° C.; thereaction is carried out for a suitable time, for example 1 to 24 hours,and for example 2 to 7 hours; step III: removing the protection group ofcompound e to provide a compound of general formula I: the reaction iscarried out in suitable water and/or an organic solvent, and the organicsolvent may be selected from a halogenated hydrocarbon (e.g.dichloromethane, chloroform, 1,2-dichloroethane or the like), a nitrile(e.g. acetonitrile or the like), an alcohol (e.g. methanol, ethanol), anether (e.g. tetrahydrofuran, dioxane), N-methylpyrrolidone,dimethylformamide, dimethylacetamide, dimethylsulfoxide and anycombination thereof, preferably water and ethanol; the reaction iscarried out under suitable acidic condition, and the acid ishydrochloric acid, sulfuric acid or trifluoroacetic acid; the reactionis carried out under suitable basic condition, and the base is sodiumhydroxide, potassium hydroxide, sodium carbonate, potassium carbonate orcesium carbonate, preferably the base is selected from sodium hydroxide;the reaction is carried out at a suitable temperature, and the reactiontemperature is preferably from −20 to 100° C., more preferably 0 to 60°C.; the reaction is carried out for a suitable time, for example 1 to 24hours.
 27. A pharmaceutical composition, comprising at least onecompound according to claim 12 or a pharmaceutically acceptable salt,ester, solvate (e.g., hydrate), stereoisomer, tautomer, prodrug thereof,or crystalline form, metabolite thereof, or a mixture of theaforementioned, and one or more pharmaceutically acceptable carriers;preferably, the pharmaceutical composition further comprises one or moreother drugs that prevent or treat a CCR2- and/or CCR5-mediated diseaseor condition, especially nonalcoholic fatty liver disease (NAFLD) or thelike; preferably, the pharmaceutical composition is in a form selectedfrom tablets, capsules, lozenges, hard candies, powder, sprays, creams,ointments, suppositories, gels, pastes, lotions, ointments, aqueoussuspensions, injectable solutions, elixirs and syrups.
 28. A method forthe prevention or treatment of a CCR2- and/or CCR5-mediated disease orcondition, comprising administering to an individual in need atherapeutically effective amount of the compound according to claim 12or a pharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned; preferably, the CCR2-and/or CCR5-mediated disease or condition is preferably nonalcoholicfatty liver disease (NAFLD) or the like.
 29. A pharmaceuticalcomposition, comprising at least one compound according to claim 16 or apharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned, and one or morepharmaceutically acceptable carriers; preferably, the pharmaceuticalcomposition further comprises one or more other drugs that prevent ortreat a CCR2- and/or CCR5-mediated disease or condition, especiallynonalcoholic fatty liver disease (NAFLD) or the like; preferably, thepharmaceutical composition is in a form selected from tablets, capsules,lozenges, hard candies, powder, sprays, creams, ointments,suppositories, gels, pastes, lotions, ointments, aqueous suspensions,injectable solutions, elixirs and syrups.
 30. A method for theprevention or treatment of a CCR2- and/or CCR5-mediated disease orcondition, comprising administering to an individual in need atherapeutically effective amount of the compound according to claim 16or a pharmaceutically acceptable salt, ester, solvate (e.g., hydrate),stereoisomer, tautomer, prodrug thereof, or crystalline form, metabolitethereof, or a mixture of the aforementioned; preferably, the CCR2-and/or CCR5-mediated disease or condition is preferably nonalcoholicfatty liver disease (NAFLD) or the like.